EMBO/soda-nel huggingface dataset:
Rab21 regulates caveolin-1-mediated endocytic trafficking to promote immature neurite pruning (2022)
Figure 2

Figure 2. Rab5 and Rab21 have distinct functions in vivo.

(A-D) Cerebral cortices at E17, electroporated with the indicated plasmids plus pCAG-EGFP at E14. (A) High magnification images of the upper intermediate zone (u-IZ) or lower intermediate zone (lo-IZ) of the cerebral cortices. White arrows indicate abnormal primary neurites. (B) High magnification images of the lower IZ of the cerebral cortices. White arrows indicate sticky neurites. (C) The ratio of locomoting neurons with more than three primary neurites in the IZ. Each score represents the mean with the individual points. Control: n = 4 brains (60 cells), Rab21-sh115: n = 3 brains (65 cells), Rab5-sh232: n = 4 brains (111 cells). (D) The box-and-whisker plot shows average leading process length of the locomoting neurons in the IZ. Control: n = 100 cells, Rab21-sh115: n = 80 cells, Rab5-sh232: n = 111 cells.

(E) Time-lapse imaging of control and Rab21-sh115-electroporated cells in cortical slices from E16 cerebral cortices, electroporated with the indicated plasmids at E14. After formation of the leading process, control neurons rapidly eliminated their immature neurites (yellow arrows), whereas the Rab21-knockdown neurons retained the immature neurites (white arrows) for long periods. Time interval of each frame is 30 min.

Data information: (C) Significance compared to control was determined by Mann-Whitney's U test (Control vs Rab21-sh232: P = 0.02771, Control vs Rab5-sh232: P = 0.2367) and Welch's t test (Control vs Rab21-sh232: P = 0.007987, Control vs Rab5-sh232: P = 0.4819). *: P<0.05, n.s.: no significant differences. (D) In the box-and-whisker plots, the central band and the upper and lower sides of the boxes indicate the median and the upper and lower quartiles. The whiskers of the depicted boxplots go from the minimum to the lower quartile and from the upper quartile to the maximum. "x" indicates the average value. Significance compared to control was determined by Mann-Whitney's U test (Control vs Rab21-sh232: P = 0.00006332, Control vs Rab5-sh232: P = 0.5213) and Welch's t test (Control vs Rab21-sh232: P = 0.00001667, Control vs Rab5-sh232: P = 0.4581). **: P<0.01, n.s.: no significant differences. Scale bars: 10 μm in (A), 2 μm in (B), 20 μm in (E).
Rab21 regulates caveolin-1-mediated endocytic trafficking to promote immature neurite pruning (2022)
Figure 3

Figure 3. Rab21 prefers to colocalize with caveolin-1 in cortical neurons.

(A-C) Primary cortical neurons from E15 cerebral cortices were transfected with pCAG-PM-mAG1 (green) and incubated for two days in vitro. Cells were immunostained with the indicated antibodies (red and blue/white). The transfected PM-mAG1 is a marker for the plasma membrane. The images are obtained with high-resolution microscopy (Nikon) and the lower panels are high magnification images near the plasma membrane. Blue alone channels are shown in black and white images. The graphs in (C) show the Pearson's correlation coefficient between PM-mAG1 and Rab21 or Caveolin-1 (left) and between caveolin-1 and Rab5 or Rab21 in whole cells (middle) or at the plasma membrane (right). Each score represents the mean of ratios with the individual points. Rab5: n = 18 cells (middle and right), Rab21: n = 31 cells (left and middle) or 20 cells (right), Caveolin-1: n = 43 cells (left).

(D-G) Immature neurons in the IZ of the cerebral cortices at E17, electroporated with pCAG-EGFP at E14. Frozen sections were immunostained with the indicated antibodies. The images are obtained with high-resolution microscopy (Nikon). Blue alone channels are shown in black and white images. The images in (F) are high magnification images of (E). The graph in (G) shows the Pearson's correlation coefficient between caveolin-1 and Rab5 or Rab21. Each score represents the mean of ratios with the individual points. Rab5: n = 38 cells, Rab21: n = 28 cells.

(H-I) HeLa cells were immunostained with the indicated antibodies. The images are obtained with high-resolution microscopy (Nikon). The graph in (I) shows the Pearson's correlation coefficient between caveolin-1 and Rab5 or Rab21. Each score represents the mean of ratios with the individual points. Rab5: n = 80 cells, Rab21: n = 62 cells.

Data information: (C) Significance was determined by Student's t test (Middle: P = 9.144E-22, Right: P = 2.309E-19). **: P<0.01. (G) Significance was determined by Welch's t test (P = 1.225E-25). **: P<0.01. (I) Significance was determined by Welch's t test (P = 3.209E-40). **: P<0.01. Scale bars: 3 μm in (upper panels in A, B, D, E), 0.5 μm in (lower panels in A, B, D, E), 0.1 μm in (F), 10 μm in (left panels in H), 1 μm in (right panels in H).
Rab21 regulates caveolin-1-mediated endocytic trafficking to promote immature neurite pruning (2022)
Figure 4

Figure 4. Rab21, but not Rab5, regulates caveolin-1-mediated trafficking pathways.

(A-F) Primary cortical neurons from E15 cerebral cortices were transfected with the indicated plasmids plus pCAG-EGFP (C and E) or pCAG-PM-mAG1 (D), incubated for two days in vitro and treated with BODIPY-LacCer (LacCer) (green) (A-B) or Alexa555-conjugated Cholera Toxin Subunit B (CTxB) (red) (C-F) for 30 minutes before fixation. White arrows in (A) indicate the perinuclear accumulation of LacCer. Blue alone channels are shown in black and white images. The graphs in (B) and (F) show the ratio of cells with perinuclear accumulation of LacCer (B) or CTxB (F), which was quantified in a blinded counting. Each score represents the mean with the individual points. B: n = 5 biological replicates (Control: 115 cells, Cav1-sh490: 127 cells, Rab21-sh115: 136 cells, Rab5-sh232: 216 cells, Rab21-sh115 + wt-Rab21: 132 cells), F: n = 4 biological replicates (Control: 64 cells, Cav1-sh490: 128 cells, Rab21-sh115: 130 cells, Rab5-sh232: 56 cells, Rab21-sh115 + wt-Rab21: 58 cells).

Data information: (B and F) Significance compared to control was determined by one-way ANOVA with post hoc Dunnett and Tukey-Kramer. **: < the critical value at 1%, n.s.: no significant differences. Scale bars: 3 μm in (A), 10 μm in (C), 2 μm in (D), 5 μm in (E).
A pseudouridine synthase module is essential for mitochondrial protein synthesis and cell viability (2016)
Figure 1

Figure 1. Mitochondrial pseudouridylation module.

A. Identification of FASTKD2 interacting proteins by AP-MS and BioID. 26 high confidence interactors detected by both techniques are listed on the right.

B. Prey specificity graph for BioID interactome of TRUB2 protein. Prey specificity was calculated as the relative enrichment of interaction of individual preys and TRUB2, compared to their interaction with 138 other baits (42 mitochondrial baits, 96 baits from other cellular compartments).
A pseudouridine synthase module is essential for mitochondrial protein synthesis and cell viability (2016)
Figure 2

Figure 2. Mitochondrial pseudouridine synthases TRUB2, RPUSD3 and RPUSD4 are necessary for OXPHOS biogenesis.

A. Immunoblot analysis of indicated proteins in control and siRNA treated cells.

B. BN-PAGE analysis of siRNA-mediated depletion shows an OXPHOS defect as revealed by subunit-specific antibodies against individual OXPHOS complexes.

C. Quantification of the levels of individual OXPHOS complexes normalized to Complex II levels. The graph represents the relative abundance of individual complexes in cells treated with the specified siRNA versus controls. The bars represent the mean ± SEM of 3-7 independent experiments. P-values were calculated using paired two-tailed t-test (* P<0.05; ** P<0.01; ***P<0.001).

D, E. Pulse-labeling mitochondrial translation experiment of the 13 mitochondria-encoded polypeptides (seven subunits of complex I [ND], three subunits of complex IV [COX], two subunits of complex V [ATP] and one subunit of complex III [cyt b]) in control and siRNA treated cells. (E) Detail of a mitochondrial pulse-labeling experiment showing a severe decrease in the translation of ATP6 and ATP8 in TRUB2 depleted cells.
A pseudouridine synthase module is essential for mitochondrial protein synthesis and cell viability (2016)
Figure 3

Figure 3. Interactions between the pseudouridine synthase module and the mitochondrial ribosome.

A, B. Identification of mitochondrial ribosomal proteins and pseudouridine synthase interacting proteins by sucrose gradient centrifugation in control cells (A) and cell treated with siRNA (B). Individual fractions were separated by SDS-PAGE and immunoblotted with the indicated antibodies. The migration of the mt-SSU (28S), the mt-LSU (39S) and the mitochondrial monosome (55S) are shown.

C. Quantification of the levels of the mt-SSU (28S), the mt-LSU (39S) and the mitochondrial monosome (55S) normalized to the SDHA levels. The graph represents the relative abundance of individual subunits in cells treated with specified siRNA versus controls. The bars represent the mean ± SEM.

D. 2D-immunoblot analysis (BN-PAGE/SDS-PAGE) of pseudouridine synthase interacting proteins. The migration (sizes in kDa) of known protein complexes in the first dimension is indicated on the top of the blot.
Mitochondrial glycerol 3-phosphate dehydrogenase promotes skeletal muscle regeneration (2018)
Figure 1

Figure 1. mGPDH regulates myoblast differentiation.

A,B qRT-PCR (A) and immunoblot (B) of mGPDH, myogenin and myosin heavy chain (MyHC) levels during C2C12 myocyte differentiation. Quantification represents the levels of indicated protein normalized to β-actin.

C Immunoblot of mGPDH, voltage-dependent anion channel (VDAC) and cytochrome c (Cyt C) levels in mitochondrial lysate during C2C12 myocyte differentiation. Quantification represents the levels of indicated protein normalized to COX IV.

D Activity assay of mGPDH at days 0 and 7 after C2C12 myocyte differentiation.

E-G Representative images of MyHC immunofluorescence (E) of C2C12 myocyte transfected with the siRNA or the overexpression plasmid for mGPDH; the fusion index (F) and the distribution of nuclei per myotube (G) were calculated at day 5 after differentiation.

H-K Immunoblot of mGPDH, myogenin and MyHC in C2C12 myocytes transfected with siRNA targeting mGPDH. Quantification (I-K) represents the levels of indicated protein normalized to β-actin at indicated day after differentiation.

L,M qRT-PCR analysis of mGPDH, myogenin and MyHC in C2C12 myocytes transfected with the siRNA or the overexpression plasmid for mGPDH at day 4 after differentiation.

Data information: Data are presented as the mean ± s.e.m. Scale bars represent 50 μm in panel E. In panels A-D and H-M, n = 3; in panels E-G, n = 15. *P < 0.05, **P < 0.01, ***P < 0.001. Unpaired t-test was used for all analyses except in panel G, where Kolmogorov-Smirnov test was used.
Mitochondrial glycerol 3-phosphate dehydrogenase promotes skeletal muscle regeneration (2018)
Figure 2

Figure 2. mGPDH is essential to skeletal muscle regeneration.

A,B qRT-PCR (A) and immunoblot (B) of mGPDH, myogenin and developmental myosin heavy chain (myh8, myl4 and myh3) in gastrocnemius (GA) muscle from C57BL/6J mice at indicated day after CTX intramuscular injection.

C Activity assay of mGPDH in GA muscle from C57BL/6J mice at day 0 and 7 after CTX injection.

D-G Representative images of the H&E staining (arrowhead, necrotic myofibers; asterisks, regenerating fibers) (D), distribution of the fibers cross-section area (CSA) (E), percentage of myofibers with central nuclei (F), and immunofluorescence staining of desmin (green) (G) in GA muscle from WT and mGPDH-/- mice at day 7 post CTX injection.

H,I Muscle weight (H) and trichrome staining (I) in GA muscle from WT and mGPDH-/- mice at day 14 post CTX injection. Quantification represents the fibrotic areas.

J,K qRT-PCR (J) and immunoblot (K) for mGPDH, myogenin and myh3 in GA muscle from WT and mGPDH-/- mice at day 7 post CTX injection.

L-Q qRT-PCR for mGPDH, myogenin and myh3 (L), H&E staining (M), distribution of the fibers CSA (N), qRT-PCR (O) and immunofluorescence staining (P) for utrophin and trichrome staining (Q) in GA muscle from mdx mice 4 weeks after AAV-mGPDH intramuscular injection.

R Exercise capacity of mdx mice 6 weeks after AAV-mGPDH tail vein injection.

Data information: Data are presented as the mean ± s.e.m. Scale bars represent 100 μm (25 μm for magnification insets) in panels D, I, M, and Q; 50 μm in panels G, P. In panels A-C, n = 3; in panels D-R, n = 6 mice per group; in panels D-F, M, and N, 3 sections were obtained per mouse. *P < 0.05, **P < 0.01, ***P < 0.001. Unpaired t-test was used for all analyses except in panels E and N, where the Kolmogorov-Smirnov test was used.
Mitochondrial glycerol 3-phosphate dehydrogenase promotes skeletal muscle regeneration (2018)
Figure 3

Figure 3. mGPDH effect occurs via the CaMKKβ/AMPK control of mitochondrial biogenesis.

A-F Mitochondrial DNA (A), nuclear-encoded OXPHOS genes (B), respirometry analysis (C) and immunoblots of mGPDH, phospho-Thr172 AMPK (p-AMPK), total AMPK (AMPK), phospho-Ser79-ACC (p-ACC), total ACC and PGC1α and corresponding quantifications represent mGPDH, p-AMPK, p-ACC and PGC1α protein levels (D-F) in C2C12 myocytes transfected with siRNA or plasmid for mGPDH 24 h after differentiation.

G-I Immunoblots of p-AMPK, p-ACC and PGC1α and corresponding quantifications represent p-AMPK, p-ACC and PGC1α protein levels (G), mitochondrial DNA (H) and nuclear-encoded OXPHOS genes combined by NDUFS8, SDHb, Uqcrc1, COX5b and ATP5a1 (I) in C2C12 myocytes transfected by mGPDH plasmid with the AMPK inhibitor compound C (CC) 24 h after differentiation.

J,K NAD+/NADH ratio (J) and immunoprecipitation analysis for PGC-1α acetyl-lysine (Ac-Lys) level (K) in C2C12 myocytes transfected with siRNA or plasmid for mGPDH 24 h after differentiation.

L-P Immunoblot of c-myc and myogenin (L) and corresponding quantifications represent c-myc and myogenin protein levels (M), representative images of MyHC immunofluorescence (N), fusion index (O) and the distribution of nuclei per myotube (P) in C2C12 myocytes transfected with mGPDH plasmid with the AMPK inhibitor CC at 24 h (L,M) or 72 h (N-P) after differentiation.

Q Immunoblots of p-AMPK, p-ACC, PGC1α and myogenin in C2C12 myocytes transfected with mGPDH plasmid with the CaMKKβ inhibitor STO-609 at 24 h after differentiation. Quantifications represent p-AMPK, p-ACC, PGC1α and myogenin protein levels.

R Immunoblots of p-AMPK and p-ACC in C2C12 myocytes transfected with mGPDH plasmid with the Ca2+ chelator BAPTA-AM at 24 h after differentiation. Quantifications represent p-AMPK and p-ACC protein levels.

Data information: Data are presented as the mean ± s.e.m. Scale bars represent 50 μm in panel N; In panels A, B, D-M, Q and R, n = 3; in panel C, n = 10; in panels N-P, n = 15. *P < 0.05, **P < 0.01, ***P < 0.001, n.s., not significant. Unpaired t-test was used in panels A-C, E, F and J; one-way ANOVA with Turkey's comparison test was used in panels G-I, M, O, Q and R; Kolmogorov-Smirnov test was used in panel P.
Mitochondrial glycerol 3-phosphate dehydrogenase promotes skeletal muscle regeneration (2018)
Figure 4

Figure 4. Rescuing mGPDH deficiency improves skeletal muscle regeneration during obesity and diabetes.

A-C Immunoblot (A,C) and IHC (B) of mGPDH and myogenin in GA muscles from obese patients (A,B) and indicated mice (C).

D qRT-PCR of mGPDH in GA muscle of indicated mice at days 0 and 3 post CTX intramuscular injection.

E-H Experimental setup (upper of E), qRT-PCR of mGPDH, myogenin and myh3 (bottom of E), H&E staining (arrowhead, necrotic myofibers; asterisks, regenerating fibers) (F), distribution of the fiber CSA (G), and percentage of myofibers with central nuclei (H) in GA muscle from AAV-mGPDH-treated HFD-fed mice at days 7 post CTX intramuscular injection.

I-M Experimental setup (upper of I, M), qRT-PCR of mGPDH, myogenin and myh3 (bottom of I), H&E staining (arrowhead, necrotic myofibers; asterisks, regenerating fibers) (J), distribution of the fibers CSA (K), percentage of myofibers with central nuclei (L) and muscle weight (bottom of M) in GA muscle from AAV-mGPDH-treated STZ-treated mice at day 7 (I-L) and 14 (M) post CTX intramuscular injection.

N Immunoblots of mGPDH, p-AMPK, p-ACC, PGC1α and myogenin for the experiment described in E.

Data information: Data are presented as the mean ± s.e.m. Scale bars represent 200 μm in panel B, 100 μm (25 μm for magnification insets) in panels F and J; In panels A and B, obese patients (n = 11) and normal subjects (n = 18); in panels C, D, n = 3 mice per group; in panels E-L and N, n = 6 mice per group; in panel M, n = 4 mice per group; in panels B, F-H, and J-L, 3 sections were obtained per mouse. *P < 0.05, **P < 0.01, ***P < 0.001. Unpaired t-test was used in panels D, E, H, I, L, and M; Wilcoxon test was used in panel B; the Kolmogorov-Smirnov test was used in panels G, K.

Expanded View Figure Legends
A KDM6A-KLF10 reinforcing feedback mechanism aggravates diabetic podocyte dysfunction (2019)
Figure 1

Figure 1. Multi-layered epigenetic controls contribute to nephrin down-regulation in immortalized mouse podocytes.

A Western blot analysis of nephrin expression in immortalized podocytes cultured in normal (5 mM) or high glucose (HG; 30 mM) or high mannitol (Mann; 5 mM glucose plus 25 mM mannitol) for 24, 48 and 72 hours. *P < 0.05 versus normal controls for the indicated time points (Wilcoxon two-sample test; n = 3).

B Analysis of the methylation status at nephrin gene promoter in podocytes cultured in normal, high glucose or high mannitol at the indicated time points. U, unmethylated-specific primers; M, methylated-specific primers. Experiments were repeated three times and a representative gel from one experiment is shown.

C Effect of pargyline hydrochloride, a pan-KDM inhibitor, on high glucose-mediated nephrin expression. Immortalized mouse podocytes were cultured in the indicated cultured media in the presence or absence of pargyline hydrochloride (1 μM) for 48 hours. Protein lysates from the treated podocytes were analyzed for nephrin expression by immunoblotting. *P < 0.05 versus normal controls, #P < 0.05 versus untreated HG-incubated cells (Wilcoxon two-sample test; n = 3).

D Effect of pargyline hydrochloride on high glucose-mediated DNA methylation at nephrin gene promoter. Genomic DNA samples from the treated podocytes in (C) were analyzed for methylation status of nephrin gene promoter by methylation-specific PCR. Experiments were repeated three times and a representative gel from one experiment is shown.

E Quantitative RT-PCR analyses of different KDMs in immortalized podocytes exposed to high glucose or high mannitol for 48 hours. *P < 0.05 versus normal controls (Wilcoxon two-sample test; n = 3).

F Western blot analysis of KDM6A expression in podocytes cultured in normal, high glucose or high mannitol conditions for 24, 48 ad 72 hours. *P < 0.05 versus normal controls for the indicated time points (Wilcoxon two-sample test; n = 3).

G Western blot analysis of H3K27me1, H3K27me2, H3K27me3 and pan-methyl H3K9 in podocytes cultured in normal, high glucose or high mannitol conditions for 9 and 24 hours. *P < 0.05 versus normal controls (Wilcoxon two-sample test; n = 3).

Data information: Data are expressed as mean ± SEM. See the exact P values for comparison tests in Appendix Table S1.
A KDM6A-KLF10 reinforcing feedback mechanism aggravates diabetic podocyte dysfunction (2019)
Figure 2

Figure 2. KDM6A contributes to down-regulation of neprhin, WT-1 and H3K27me3 in podocytes.

A Effect of KDM6A knockdown on the expression of nephrin and WT-1 in immortalized mouse podocytes cultured under high glucose conditions. Western blot analyses were performed using protein lysates of podocytes that were transfected with scrambled siRNAs or KDM6A-specific siRNAs, and cultured in high glucose (HG) or high mannitol (Mann) for 48 hours. *P < 0.05 versus normal controls, #P < 0.05 versus control siRNA with HG incubation (Parametric ANOVA and a Bonferroni post hoc test; n = 3).

B Effect of KDM6A overexpression on expression levels of nephrin and WT-1. Immortalized podocytes were transfected with the KDM6A expression plasmid or the empty vector control for 48 hours, and the expression levels of KDM6A, nephrin and WT-1 in these transfected podoytes were analyzed by immunoblotting. *P < 0.05 versus vector controls (Parametric ANOVA and a Bonferroni post hoc test; n = 3).

C Changes in H3K27me3 levels in podocytes after treatment with high glucose, the combination of high glucose and KDM6A knockdown, or KDM6A overexpression for 48 hours. *P < 0.05 versus normal controls, #P < 0.05 versus control siRNA with HG incubation (Parametric ANOVA and a Bonferroni post hoc test; n = 3).

D Changes in DNA methylation at nephrin gene promoter by high glucose, by the combination of high glucose plus KDM6A knockdown, or by KDM6A overexpression in immortalized podocytes. U, unmethylated-specific primers; M, methylated-specific primers. Experiments were repeated three times and a representative gel from one experiment is shown.

E Effect of GSK-J4 (40 μM), a specific KDM6A inhibitor, on high glucose-mediated changes in levels of nephrin, WT-1 and H3K27me3. *P < 0.05 versus normal controls, #P < 0.05 versus untreated HG-incubated cells (Parametric ANOVA and a Bonferroni post hoc test; n = 3).

F Changes in DNA methylation at nephrin gene promoter by GSK-J4 (40 μM) in immortalized podocytes. U, unmethylated-specific primers; M, methylated-specific primers. Experiments were repeated three times and a representative gel from one experiment is shown.

Data information: Data are expressed as mean ± SEM. See the exact P values for comparison tests in Appendix Table S2.
A KDM6A-KLF10 reinforcing feedback mechanism aggravates diabetic podocyte dysfunction (2019)
Figure 3

Figure 3. KDM6A promotes podocyte and kidney dysfunction in diabetic mice.

A Relative mRNA levels of KDM6A, nephrin and WT-1, normalized to β-actin, expressed in kidney glomeruli of normal and diabetic mice at 4, 8 and 12 weeks after diabetic induction. *Significant differences (P < 0.05) compared with normal controls (Wilcoxon two-sample test; n =8 each).

B Western blot analysis of nephrin, KDM6A and H3K27me3 in kidney glomeruli of normal and diabetic mice at different time points. *P < 0.05 versus normal controls for the indicated time points (Wilcoxon two-sample test; n = 3).

C Changes in body weights and levels of blood glucose in normal, diabetic and GSK-J4-treated diabetic mice. As noted, GSK-J4 treatment did not significantly affect body weights or blood glucose levels in diabetic mice during the experimental period of 12 weeks (Wilcoxon two-sample test; n = 8).

D Levels of urinary protein excretion, weights of kidney, and levels of HbA1c in normal, diabetic and GSK-J4-treated diabetic mice. Urinary total protein excretion, kidney weight and levels of HbA1c were measured at 12 weeks after diabetic induction. The mean relative kidney weight (%) shown in the study is determined as the percent of kidneys out of total body weight, and the HbA1c level is defined as the ratio of HbA1c to the total hemoglobin (% HbA1c; DCCT unit). *P < 0.05 versus normal controls, #P < 0.05 versus untreated diabetic mice (Parametric ANOVA and a Bonferroni post hoc test; n = 8).

E Western blot analysis of KDM6A, nephrin, WT-1 and H3K27me3 expressed in kidney glomeruli of normal, diabetic and GSK-J4-treated diabetic mice at 12 weeks after diabetic induction. *P < 0.05 versus normal controls, #P < 0.05 versus untreated diabetic mice (Parametric ANOVA and a Bonferroni post hoc test; n = 3).

F Immunofluorescence analysis of KDM6A, nephrin and WT-1 in kidney sections from normal, diabetic, and GSK-J4-treated diabetic mice. Green: nephrin or WT-1; Red: KDM6A; Blue: DAPI. Scale bars, 20 μm. *P < 0.05 versus normal controls, #P < 0.05 versus untreated diabetic mice (Parametric ANOVA and a Bonferroni post hoc test; n = 3).

G Immunofluorescence staining of F-actin and KDM6A in primary podocytes isolated from normal, diabetic, and GST-J4-treated diabetic mice. Green: F-actin; Red: KDM6A; Blue: DAPI. Scale bars, 20 μm. Presented experiments were performed at least three times independently.

Data information: Data are expressed as mean ± SEM. See the exact P values for comparison tests in Appendix Table S3.