EMBO/soda-nel huggingface dataset:
AKT2 reduces IFNβ1 production to modulate anti-viral responses and systemic lupus erythematosus (2021)
Figure 1

Figure 1. Akt2 expression is negatively correlated with IFNβ1 production

A The mRNA value of AKT2 was analyzed in the GEO Profiles from the liver explant of Hepatitis B virus (HBV)-associated acute liver failure (ALF) patients (GDS4387/225471_s_at), and from bronchial epithelial cells with pandemic and seasonal H1N1 influenza virus infections in vitro (GDS4855/203808_at). normal, n=10; HBV, n=17; control, n=3; influenza A, n=9.

B The mRNA expression of Akt2 in the brain homogenates of WT mice with JEV injection (5.0×106 PFU/g, i.v.) or in the lung of WT mice with H7N9 (105.5 EID50, i.n.) and PR8 infection (10 LD50, i.n.) were detected by quantitative reverse transcription-PCR (qRT-PCR). Mock, n=4; JEV, n=12; PBS, n=6; H7N9, n=4; PR8, n=8.

C The qRT-PCR analysis of Akt2 mRNA in PEMs stimulated with lipo-poly(I:C) (1 μg/mL), lipo-poly(A:T) (1 μg/mL), lipo-ISD (3 μg/mL) or HSV-1 (MOI, 1) for 6 hours. n=3, respectively.

D The mRNA of Akt2 (red line) and Ifnb1 (black line) were analyzed by qRT-PC in the mock and VSV (MOI, 1) or LM (MOI, 1)-stimulated PEMs or BMDMs at the indicated time. Mock and VSV, n=4; mock and LM, n≥3.

E The mRNA levels of Akt2 in WT (n=3) and Ifnar1 KO (n=3) PEMs stimulated with VSV and lipo-ISD for 6 hours were detected by qRT-PCR.

F PEMs were pretreated with the indicate inhibitors for 2 hours and stimulated with recombined of IFNβ1 (1 μg/mL) for another 3 hours, followed by qRT-PCR for detection of the Akt2 mRNA levels. DMSO, n=2; inhibitors, n=3.

G-H PEMs were pretreated with DMSO or AKT2 inhibitor CCT128930 (10 μM) (G) for 2 hours, or knocked down of Akt2 with siAkt2-1 (20 nM) for 48 hours (H), then the mRNA level of Ifnb1 was measured by qRT-PCR after lipo-poly(I:C), lipo-poly(A:T), lipo-ISD, VSV, or HSV-1 stimulation for another 6 hours. n=3, respectively.

Data information: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 and ns, not significant (P > 0.05); using unpaired t-test (A, B left panel), or one-way ANOVA test (B right panel, C and F), or two-way ANOVA test (E, G and H). Data are from two (F) or at least three independent biological replicates (C-E, G, H). Error bars (A and B, mean ± SD; C-H, mean ± SEM). Source data are available online for this figure.
AKT2 reduces IFNβ1 production to modulate anti-viral responses and systemic lupus erythematosus (2021)
Figure 2

Figure 2. AKT2 kinase activity is indispensable to attenuate I-IFN production in macrophages and in zebrafish larvae

A-B qRT-PCR detection for the mRNA expression of Ifnb1 (A), Ifna4, Cxcl10 and Ccl5 (B) in WT (n=3) and Akt2 KO (n=3) PEMs stimulated with lipo-poly(I:C), VSV, lipo-poly(A:T), lipo-ISD or HSV-1 for 6 hours.

C The expression of Ifnb1 mRNA level by qRT-PCR in WT (n=3) and Akt2 KO (n=3) PEMs treated with poly(I:C) (10 μg/mL) for 6 hours (left panel) or LPS (1 μg/mL) for 2 hours (right panel).

D The expression of Ifnb1 in WT (n=3) and Akt2 KO (n=3) primary-MEFs with VSV treatment (MOI, 0.1) for 6 hours was measured by qRT-PCR (left panel). After VSV treatment for 3 hours, the culture suspension was discarded and wash 3 times with PBS, then primary-MEFs was cultured with fresh medium for another 15 hours for analysis of the VSV-infected (GFP) primary-MEFs by fluorescence microscope (middle panel, representative images) and by FACS assay (right panel). Bar, 100 μm.

E MEF (left panel, n=3) and HEK293T cells (right panel, n=3) were transfected with AKT2 for 24 hours and stimulated with VSV for 6 hours, then cells were harvested for detection the mRNA expression of Ifnb1 by qRT-PCR.

F IFNβ1 luciferase assays of HEK293T cells with transfection of AKT2/AKT2-T309A/S474A and TBK1 (n=3) or infection of VSV (n=3). Protein expression levels are shown in Figure EV2F.

G Zebrafish larva at 48 hour after fertilization were micro-injected GFP-fused VSV (1 × 103 PFU/larvae) for 18 hours, then the representative images of VSV-infected zebrafish larva were collected by fluorescence microscope. The infected area (GFP) and macrophages (Red) are indicated by arrows. Bars, 200 μm.

H Zebrafish larvae were overexpressed the indicated protein for 48 hours and challenged with VSV for another 6 hours, then the mRNA levels of ifn1 in zebrafish larvae were measured by qRT-PCR (left panel). Every dot represents three zebrafish embryos. Horizontal square bracket shows the statistical analysis of the comparison with 'PBS mock', the rest shows the comparison with 'PBS VSV'. PBS mock, n=5; PBS VSV, n=13; AKT2 VSV, n=7; AKT2-T309A/S474A VSV, n=9. H&E staining (middle panel) and survival rates (Kaplan-Meier curve) (right panel) were collected from zebrafish larvae after VSV micro-injection for 18 hours or longer. The arrows indicated the VSV-infected eye and skeletal muscle in zebrafish larvae. Bars, 100 μm.

Data information: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 and ns, not significant (P > 0.05); using a one-way ANOVA test (E, F, H left panel), or two-way ANOVA test (A-C, D left and right panel), or log-rank (Mantel-Cox) test (H right panel). Data are from three independent experiments (A-C, D left and right panel, E, F) or representative of three independent biological replicates (D middle panel, G, H). Error bars (A-F, mean ± SEM; H, mean ± SD). Source data are available online for this figure.
AKT2 reduces IFNβ1 production to modulate anti-viral responses and systemic lupus erythematosus (2021)
Figure 3

Figure 3. AKT2 restrains IRF3 nuclear translocation via 14-3-3ε

A and B IFNβ1 luciferase assays (upper panels) in HEK293T (A) and HEK293T-IRF3 KO (B) cells transfected with the indicated plasmids or infected with VSV. Immunoblot analysis showed the indicated constructs in HEK293T cells (bottom panels). n=3, respectively.

C The mRNA levels of Ifnb1 were measured by qRT-PCR in WT (n=3) and Akt2 KO (n=3) PEMs with Irf3 knockdown for 48 hours and VSV treatment for 6 hours.

D WT and Akt2 KO PEMs were infected with or without VSV for 6 hours. Then, cell lysates of WT or Akt2 KO PEMs were prepared for immunoprecipitation using anti-AKT2 antibody, followed by immunoblotting using anti-IRF3 antibody (left panel). Alternatively, cell lysates were prepared for immunoprecipitation using anti-IRF3 antibody, followed by immunoblotting using anti-AKT2 antibody (right panel) to detect the endogenous interaction between AKT2 and IRF3.

E Immunoblot analysis of GST-AKT2 and His-IRF3 interaction in a GST pull-down assay.

F Immunoassay of IRF3 in dimer or monomer form by native-gel and p-IRF3 (Ser396), p-TBK1 (Ser172), AKT2 and ACTIN by SDS-gel in WT and Akt2 KO PEMs with VSV stimulation for 6 hours.

G Immunofluorescent microscopic imaging for (left panel) and statistics analysis (right panel) for IRF3 nuclear translocation in WT and Akt2 KO PEMs at 6 hour post-VSV infection. IRF3 (red), Nuclei (Hoechst, green). The white arrows indicate the nuclei with IRF3 translocation. Mock, n=2; VSV, n=3. Bar, 20 μm.

H Immunoassay of IRF3, AKT2, LaminB1 and GAPDH in the nuclear and cytoplasmic fractions from the WT and Akt2 KO PEMs after VSV treatment for 6 hours. Tubulin and LaminB1 were used as cytoplasmic and nucleic protein loading control respectively.

I Immunoassay of IRF3, SP1 and Tubulin in the nuclear and cytoplasmic fractions of the HEK293T cells overexpressed with indicated constructs for 24 hours and stimulated with VSV for 6 hours. Tubulin and SP1 served as cytoplasmic and nucleic protein loading control respectively.

J IFNβ1 luciferases assays were performed in HEK293T cells transfected with AKT2 and 14-3-3ε followed by VSV treatment. n=3, respectively.

K qRT-PCR analysis for the Ifnb1 mRNA levels in VSV-stimulated WT and Akt2 KO PEMs with 14-3-3ε siRNA pretreatment. n=3, respectively.

Data information: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 and ns, not significant (P > 0.05); using a one-way ANOVA (A, B, J), or two-way ANOVA test (C, G right panel, K). Data are from three independent experiments (A-C, J and K), or representative of two or three independent biological replicates (D-I). Error bars (A-C, G, J and K, mean ± SEM). Source data are available online for this figure.
AKT2 reduces IFNβ1 production to modulate anti-viral responses and systemic lupus erythematosus (2021)
Figure 4

Figure 4. AKT2 phosphorylates IRF3 at Thr207 and blocks IRF3 activation

A Immunoblot analysis of IRF3 in the cytoplasmic and nucleic fractions of HEK293T-IRF3 KO cells with overexpression of IRF3 and AKT2 or AKT2-S309A/T474A.

B Immunoblot analysis of in vitro kinase assay by the phosphorylation-gel. HA-AKT2 was immunoprecipitated from HEK293T cells by anti-HA/IgG antibody and Protein-G beads, and His-IRF3 was purified from E.coli cells by anti-His agarose beads. His-IRF3 alone and anti-IgG-beads added with His-IRF3 served as control. The arrow indicated the phosphorylated IRF3.

C IFNβ1 luciferase assays and immunoblot analysis showed the HEK293T-IRF3 KO cells with overexpression of the indicated plasmids for 24 hours and VSV infection for 6 hours. n=3, respectively.

D Immunofluorescence microscopy (top panel) of AKT2-, Flag-IRF3- or Flag-IRF3-T207A-overexpressed MEF cells after poly(A:T) stimulation for 6 hours. Flag (IRF3, red) and Hoechst (nuclei, blue). Immunoassay of nuclear-cytoplasm extractions (bottom panel) from HEK293T-IRF3 KO cells with overexpression of the indicated plasmids for 24 hours followed by VSV infection for 6 hours. GAPDH and SP1 were used as cytoplasmic and nucleic protein loading control respectively. Bar, 20 μm.

E HEK293T-IRF3 KO cells were overexpressed with indicated plasmids, and the cell lysate was conducted to immunoblot analysis by phosphorylation-gel for the assay of phosphorylated IRF3 (the upper band) by anti-Flag antibody.

F Survival rates (left panel) until 72 hours and H&E staining (right panel) at 18 hours of the zebrafish larvae with indicated protein expression and VSV challenge. The arrows indicated the VSV-infected eye and skeletal muscle in zebrafish larvae. Bars, 100 μm.

G qRT-PCR analysis of the VSV copies in overexpressed zebrafish embryos with the VSV challenge at 18 hours later. Every dot represents three zebrafish embryos. Horizontal vertical square bracket shows the statistical analysis of comparison with 'PBS mock', the rest shows the comparison with 'PBS VSV'. PBS mock, n=15; PBS VSV, n=9; IRF3 VSV, n=16; IRF3-T207A VSV, n=11; AKT2 VSV, n=14; AKT2-T309A/S474A VSV, n=9; IRF3 + AKT2 VSV, n=9; IRF3 + AKT2-T309A/S474A VSV, n=13; IRF3-T207A + AKT2 VSV, n=10.

H The survival rates of over-expressed zebrafish larvae as long as 72 hours after the VSV challenge.

Data information: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 and ns, not significant (P > 0.05); using a one-way ANOVA test (G), or two-way ANOVA test (C), or log-rank (Mantel-Cox) test (F left panel and H). Data are from at least three independent experiments (C), or representative of three independent experiments (A, B, D-H). Error bars (C, mean ± SEM; G, mean ± SD). Source data are available online for this figure.
AKT2 reduces IFNβ1 production to modulate anti-viral responses and systemic lupus erythematosus (2021)
Figure 5

Figure 5. Targeting AKT2 enhances antiviral defense in mice

A FACS analysis of the percentage of macrophages (CD11b+ F4/80+ cells) in WT and Akt2 KO BM and spleen.

B Survival rates of WT and Akt2 KO mice after lethal dose injection of VSV during 120 hours (1 × 107 PFU/g, i.v.).

C qRT-PCR analysis of the Ifnb1 mRNA expression in the liver (n=8), spleen (n=7) and lung (n=7) of WT or Akt2 KO mice 6 hours after the injection of VSV (1 × 106 PFU/g, i.v.).

D IFNβ1 concentrations from WT (n=8) and Akt2 KO (n=7) serum were detected by ELISA 3 hours after VSV infection (1 × 106 PFU/g, i.v.).

E WT and Akt2 KO mice were injected with VSV for 24 hours (1 × 106 PFU/g, i.v.). Representative images of the VSV infected cells (GFP) were collected by fluorescence microscope (left panel) and the VSV copies (WT, n=6; Akt2 KO n=7) were measured by qRT-PCR (right panel) in the liver of mice. Bar, 100 μm.

F-H Bone marrow cells from WT and Akt2 KO mice (CD45.2) were adoptively transferred to lethally irradiated CD45.1 mice and infected with VSV. Survival rates (F left panel), serum IFNβ1 concentrations (F right panel, n=5), VSV copies (G, n=9) and representative images (H) were analyzed as Fig 5B, 5D and 5E, respectively. Bar, 100 μm.

I Survival rates of WT mice that were intraperitoneal injected with CCT128930 (20 mg/kg) twice every other day before VSV infection (1 × 107 PFU/g, i.v.).

Data information: *P < 0.05, **P < 0.01; using a paired t-test (C and F right panel), or unpaired t-test (D, E right panel and G) or log-rank (Mantel-Cox) test (B, F left panel and I). Data are representative of three independent experiments respectively (A-I), in which the different groups of mice were sacrificed and quantified for indicated experimental purposes (C-E, F right panel, G). Error bars (D, E right panel and G, mean ± SD). Source data are available online for this figure.
AKT2 reduces IFNβ1 production to modulate anti-viral responses and systemic lupus erythematosus (2021)
Figure 6

Figure 6. Akt2 is associated with the pathology of SLE

A Representative images of lungs and spleens from WT and Akt2 KO mice treated with TMPD (500μL per mouse) for 2 weeks, and the corresponding H&E-staining of the tissues. Bar, 200 μm.

B-C qRT-PCR analysis of the relative Ifnb1 and Ifna4 mRNA expression levels in PICs (n≥5) and PBMCs (n≥4) (B), and immunoblot analysis of IRF3, LaminB1 and GAPDH in the PIC nuclear and cytoplasmic fractions (C) from WT and Akt2 KO mice treated with TMPD for 2 weeks. GAPDH and LaminB1 were used as cytoplasmic and nucleic protein loading control respectively.

D-E Representative images of lipogranulomas in peritoneal cavity (D), H&E-staining (E left panel) and immunofluorescence of IgG deposits (E right panel) in the kidney sections were collected from WT and Akt2 KO bone marrow-reconstituted CD45.1 mice 12 weeks after TMPD treatment. Bar, 50 μm.

F-H Quantitative analysis of serum anti-dsDNA IgG antibody by ELISA (F, n≥4), mRNA expression levels of PIC Ifnb1 and Ifna4 by qRT-PCR (G, n≥4) and FACS analysis of the percentages of B220+ and CD19+MHCⅡ+ cells in PBMC (H, n≥4) from WT and Akt2 KO bone marrow-reconstituted CD45.1 mice 12 weeks after TMPD treatment.

I qRT-PCR analysis of the relative mRNA expression levels of AKT2 in PBMC isolated from SLE patients (n=9) or healthy donors (n=9).

J. qRT-PCR analysis of the relative mRNA expression levels of AKT2 (left panel) and IFNA (middle panel) or IFNβ1 (right panel) in CD14 positive monocytes isolated from SLE patients (n=9) or healthy donors (n=9).

K Immunofluorescence of IRF3 (Red), AKT2 (Green), DAPI (Blue, nucleus) in CD14+ cells from SLE patients followed by analysis of the cells about mean amount of AKT2 in this cell as well as mean amount of IRF3 in the same nucleus with Image J software. Cells (n=39) were collected 3 patients. Bar, 2 μm.

Data information: *P < 0.05, **P < 0.01 and ****P < 0.0001; using unpaired t-test (B, G-I, J left panel), or two-way ANOVA test (F), or correlation analyses (J middle and right panels, K right panel). Data are pooled from different individuals (I, J, K right panel) or representative of three independent experiments (A-H, K left panel). Error bars (B, F-J, mean ± SD). Source data are available online for this figure.

Expanded View Figures
Small molecule inhibitors of the Dishevelled-CXXC5 interaction are new drug candidates for bone anabolic osteoporosis therapy (2016)
Figure 1

Figure 1. Identification and characterization of small molecules competing Dvl-CXXC5 binding in vitro.

A. A scheme for in vitro screening method of small molecules competing Dvl-CXXC5 binding. Briefly, purified Dvl PDZ domain was attached to the polystyrene surface of each well of 96-well plates. Then, PolyR-DBM (ployarginine conjugated Dvl binding motif tagged with FITC) (Kim et al, 2015) was added to each well and allowed to bind to the Dvl PDZ domain; 10 µM small molecule compound was added to each well, and the compounds competing with Dvl PZD-PolyR-DBM binding were measured by a microplate reader to screen the compounds reducing fluorescence signal.

B . Screening results of compounds competing Dvl-CXXC5 interaction. Each 2 µM of the compounds was used for competing as described in Fig 1A. [n=3]

C, D. Primary calvaria cells were treated with 2 µM of each compound for 2 days. The cells were subjected to immunofluorescence analyses to visualize β-catenin (C; green). The relative numbers of β-catenin-positive nuclei were counted (D). [n=3]

E, F . The calvariae from 4-day-old mice were cultured ex vivo for 7 days with 2 µM of each compound. Representative calvaria sections were visualized by H&E staining (E). The calvaria thicknesses were measured from the images using Image Pro software (F). [n=3]

Data Information: For B, D, and F, the data are the mean ± s.d. (error bars), and significance was assessed using unpaired Student’s t-test.
Small molecule inhibitors of the Dishevelled-CXXC5 interaction are new drug candidates for bone anabolic osteoporosis therapy (2016)
Figure 2

Figure 2. DBM-mimetic binding of KY-02061 on the Dvl PDZ domain.

A . The chemical structure of KY-02061.

B . A competition curve for the Dvl-CXXC5 interaction by KY-02061.

C-E. NMR titration analyses for Dvl PZD domain with KY-02061. 1H-15N-HSQC analyses were performed to analyze the interaction of 15N-labelled Dvl-PDZ domain with KY-02061. The 1H-15N-HSQC spectrum of different molar ratios (Dvl PDZ domain:KY-02061) are displayed as red (1:0), orange (1:10), purple (1:20), cyan (1:40), green (1:60) and blue (1:80) (C, residues with meaningful chemical shift change are indicated by arrows). Plot of chemical shift changes (Δδ) as a function of residue number in molecular ratio 1:80 (D, a red-colored line indicates the line for Δδ=0.05). The residues with Δδ greater than 0.05 are visualized as a stick model on the ribbon representation of the Dvl PDZ domain structure (E).

F. Molecular docking of Dvl binding motif (DBM) or KY-02061 to Dvl PDZ domain were analyzed by in silico experiments. The superimposed structure of DBM (green) and KY-02061 (yellow) on the surface of Dvl PDZ domain (gray) was visualized.
Small molecule inhibitors of the Dishevelled-CXXC5 interaction are new drug candidates for bone anabolic osteoporosis therapy (2016)
Figure 3

Figure 3. Effect of KY-02061 on Wnt/β-catenin pathway, osteoblast differentiation, and ex vivo-cultured calvaria growth.

A . MC3T3E1 cells were transfected with pTOPFLASH together with pCMV-β-gal. After 24 h, the cells were treated with DMSO, KY-02061 in DMSO, or 5 µM PolyR-DBM for 2 days. The luciferase activities of whole cell lysates were measured and normalized with β-galactosidase activities. [n=3]

B . Primary calvaria cells were isolated from the calvariae of 4-day-old mice (Manton et al, 2007), and treated with DMSO or KY-02061 in DMSO for 4 days. ALP activity levels were visualized by ALP staining.

C. Primary calvaria cells were treated with DMSO or 5 µM KY-02061 in DMSO for 14 days. The cells were subjected to immunofluorescence analyses to visualize Runx2 (green) and β-catenin (red). The cell nuclei were counterstained with DAPI (blue).

D, E. Calvariae from 4-day-old mice were cultured ex vivo for 7 days with KY-02061 in DMSO (D). The calvaria thicknesses were measured from the stained sections using Image Pro software (E). [n=3]

Data Information: For A and E, the data are the mean ± s.d. (error bars), and significance was assessed using unpaired Student’s t-test.
Small molecule inhibitors of the Dishevelled-CXXC5 interaction are new drug candidates for bone anabolic osteoporosis therapy (2016)
Figure 4

Figure 4. Binding of KY-02327, a KY-02061 analog, on Dvl PDZ domain.

A. The chemical structure of KY-02327.

B. A competition curve for the Dvl-CXXC5 interaction by KY-02327.

C. Fluorescence quenching plot of Dvl PDZ domain upon addition of different amounts of KY-02327. From top to bottom final moral ratio between Dvl PDZ and KY-02327 was, 1:0, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20 and 1:25, respectively.

D-F. 1H-15N-HSQC analyses were performed to analyze the interaction of 15N-labelled Dvl-PDZ domain with KY-02327. The 1H-15N-HSQC spectrum of different molar ratios (Dvl PDZ domain:KY-02327) are displayed as red (1:0), yellow (1:5), green (1:10), and magenta (1:20) (D, residues with meaningful chemical shift change are indicated by arrows). Plot of chemical shift changes (Δδ) as a function of residue number in molecular ratio 1:20 (E, a red-colored line indicates the line for Δδ=0.05). The residues with Δδ greater than 0.05 are visualized as a stick model on the ribbon representation of the Dvl PDZ domain structure (F).

G. Molecular docking of KY-02061 or KY-02327 to Dvl PDZ domain were analyzed by in silico experiments. The superimposed structure of KY-02061 (yellow) and KY-02327 (cyan) on the surface of Dvl PDZ domain (gray) was visualized.
Small molecule inhibitors of the Dishevelled-CXXC5 interaction are new drug candidates for bone anabolic osteoporosis therapy (2016)
Figure 5

Figure 5. Effects of KY-02327 on Wnt/β-catenin pathway and osteoblast differentiation in osteoblasts.

A . MC3T3E1 cells were transfected with pTOPFLASH together with pCMV-β-gal. After 24 h, the cells were treated with indicated dose of KY-02327 for 2 days. The luciferase activities of whole cell lysates were measured and normalized with β-galactosidase activities. [n=3]

B-C . MC3T3E1 cells were treated with indicated concentrations of KY-02327 for 2 days. β-Catenin and α-tubulin were detected by immunoblot (B) or immunofluorescence (C) analyses. Nuclei were counterstained with DAPI (C, blue).

D. MC3T3E1 cells were treated with indicated concentrations of KY-02327 for 14 days. The mRNA level of collagen 1a (Col1a) was measured by quantitative real-time (qRT)-PCR. [n=3]

E. MC3T3E1 cells were treated with indicated concentrations of KY-02327 for 21 days. The mRNA level of osteocalcin (OCN) was measured by qRT-PCR. [n=3]

Data Information: For A, D, and E, the data are the mean ± s.d. (error bars), and significance was assessed using unpaired Student’s t-test.
H19 lncRNA alters stromal cell growth via IGF signaling in the endometrium of women with endometriosis. (2015)
Figure 1

The expressions of H19 and Igf1r are significantly decreased in the eutopic endometrium of women with endometriosis compared to those without endometriosis

A–F (A, B, D–F) RNA levels determined by RT–qPCR. Results are presented as mean ± SD,n = 10. P-values are indicated on the top of each plot. (C) Spearman correlation suggests an in vivo positive correlation between the expressions of H19 and Igf1r in a statistically significant manner. Spearman correlation coefficient, P-values, and sample numbers are marked on the top left of the plot.
H19 lncRNA alters stromal cell growth via IGF signaling in the endometrium of women with endometriosis. (2015)
Figure 2

H19 promotes Igf1r expression in endometrial stromal cells by inhibiting let-7

A–D Endometrial stromal cells from patients #166 and #80 were each transfected with a mixture of siCon (control siRNA) and microRNA inhibitor control (iCon), siH19 and iCon, or siH19 and iLet7 (let-7-specific inhibitor). RNA and proteins were analyzed at 48 h post-transfection. Combined results from the two patient cells are presented. Western blot gels from #166 cells are shown in (C). Quantitation of Western blots combining both patient cells are shown in (D).

E–H Endometrial stromal cells from patients #98 and #212S were each transfected with empty vector or pH19. RNA and proteins were analyzed 48 h post-transfection. Combined results from the two patient cells are presented. Western blot gels from #98 cells are shown in (G). Quantitation of Western blots combining both patient cells are shown in (H).

Data information: Results are presented as mean ± SD, n = 6. P-values are indicated on the top of each plot.