EMBO/soda-nel huggingface dataset:
Repression of Irs2 by let-7 miRNAs is essential for homeostasis of the telencephalic neuroepithelium (2020)
Figure 8

Figure 8. Irs2 and let-7 regulate progenitor cell proliferation in human cerebral organoids.

A - Schema of experimental design. At 40 days in culture, human cerebral organoids were electroporated with one of the indicated DNA plasmid combinations, followed by 7 additional days in culture prior to analysis.

B-G - Stains and quantifications of human cerebral organoids electroporated with the indicated construct combinations, to reveal the frequency of electroporated cells remaining as progenitors (%GFP+Ki67+/GFP+). Histograms represent mean ± SEM; symbols in plots indicate values for individual organoids; X2-test, *P < 0.05. N = 8-11 replicates per group. Dashed lines indicate ventricular surface, solid arrowheads indicate Ki67+/GFP+ cells, open arrowheads indicate Ki67-/GFP+ cells. Scale bars, 50 µm.

H - Schematic drawing of conclusions from this study. Let-7 expressed in the early telencephalic neuroepithelium represses the expression of p53 and Irs2, which maintains the cellular homeostasis and epithelial integrity. Loss of let-7, as in Dicer1 mutants, de-represses expression of Irs2 and of p53 pathway components, which further increase Irs2 levels. High Irs2 enhances progenitor cell proliferation and loss of apical adhesion, altering neuroepithelial homeostasis and integrity, and leading to the formation of hyperproliferative rosettes.

Expanded View Figure legends
Autophagy regulates lipid metabolism (2009)
figf1

a, TG levels in hepatocytes untreated (None) or treated with 3-methyladenine (3MA) and cultured in regular medium (RM) or oleate (OL) (*P  0.02, **P  0.002, n = 3). b, TG levels in vector-infected (VEC) and siAtg5 cells in RM, OL or in MCDM (*P  0.001, n = 5). OL values are in mM. c, TG levels in wild-type (WT) or Atg5 knockout mice embryonic fibroblasts (Atg5-/-) (*P  0.01 or **P  0.0001, n = 5). df, Cells from a and b stained with BODIPY 493/503 (d), oil red O (e) or visualized by electron microscopy (f). Right: quantifications of LD number and size (*P  0.01, **P  0.001 with untreated wild-type cells; #P  0.01, # #P  0.001 with cells in RM; n = 46). Nuclei are highlighted with 4,6-diamidino-2-phenylindole (DAPI; d). Error bars, s.e.m.
Autophagy regulates lipid metabolism (2009)
figf2

a, b, VEC and siAtg5 cells cultured with oleate (OL) or in MCDM were examined for their rates of TG synthesis (a) and &bgr;-oxidation (b) as compared to cells in regular medium alone (*P  0.03, **P  0.004 with VEC cells in the same medium, n = 34). c, d, Rates of TG decay in OL (c) and MCDM (d) (*P  0.05, **P  0.01, ***P  0.001 as compared to VEC cells, n = 37). e, TG levels in wild-type cells treated with dimethyl sulphoxide vehicle (DMSO), 3-methyladenine (3MA) or diethylumbelliferyl phosphate (DEUP) (*P  0.00001 with DMSO-treated cells, #P  0.003 with 3MA-treated cells, n = 6). Error bars, s.e.m. f, Co-localization of BODIPY 493/503 (green) with LAMP1 (red) or LC3 (red) in hepatocytes in MCDM. g, High-magnification regions of hepatocytes in MCDM alone (none) or treated with vinblastine and stained as labelled. Arrows indicate co-localization.
Autophagy regulates lipid metabolism (2009)
figf3

a, Electron micrographs of cultured hepatocytes. dm, double-membrane cytosolic vesicles. Arrows indicate membranes in LDs. Insets show the double membrane in nucleus (N), lipid-containing vesicles (L) and mitochondria (mit), but not in LDs. b, Mouse liver LC3 immunogold. Insets show higher magnification. Arrowheads indicate gold particles (black) and LC3-labelled bilayer membranes (white). c, Percentages of autophagic vacuoles (AVs) containing only lipid (Lipids, *), other cargo (Other) or mixed cargo (Mixed) in cells treated as in b (*P  0.01, **P  0.001 with cells in RM, n = 46). Left: representative examples. Error bars, s.e.m.
Autophagy regulates lipid metabolism (2009)
figf4

a, Immunoblots of cellular homogenates (Hom) and LDs from fed (F) or 24-h starved (S) mice. I&kgr;B, inhibitor of the nuclear factor of kappa light polypeptide gene enhancer; GPDH, glyceraldehyde-3-phosphate dehydrogenase. b, Percentages of autophagic vacuoles (AVs) containing only lipid, other cargo or mixed cargo calculated from samples processed as in Supplementary Fig. 16b (*P  0.001, **P  0.0001, n = 46). c, Immunoblots of Hom, AV and lysosomes (Lys). L-1, LAMP1. d, Immunoblots of liver Hom and LDs from HFD-fed mice. e, Liver homogenate immunoblots. PDI, protein disulphide isomerase. f, Total hepatic TG and cholesterol content (*P  0.01, **P  0.00001, n = 817). g, Hepatic TG concentration (*P  0.05, n = 3). h, Percentage of cellular cholesterol in lysosomes (*P  0.02, n = 4). i, Immunoblots of homogenates and isolated LDs. Error bars, s.e.m.
A hit-and-run heat shock factor governs sustained histone methylation and transcriptional stress memory (2015)
Figure 1

Figure 1: HS induces sustained H3K4me3 and H3K4me2 methylation at HSP22.0, but not at HSP70.

Dynamics of histone modifications after an acclimatizing HS (ACC) or no HS (NHS) in Col-0 wild type at HSP22.0 (A) and HSP70 (B). Seedlings were subjected to ACC or a control treatment (NHS) 4 d after germination. At the indicated time points after the treatments, ChIP-qPCR was performed with antibodies against H3K9ac, H3K4me3, H3K4me2 and H3. Schematics show positions of regions analyzed. Amplicon positions relative to TSS are HSP22.0: 1, -2570 bp; 2, +235 bp. HSP70: 1, 4192 bp downstream of the 3’UTR; 2, +47 bp. Data shown are averages over three biological replicates. Amplification values were normalized to input, H3 and 4 h NHS region 2. The bottom panel shows the H3 signal normalized to input and 4 h NHS region 2. Squares and triangles within bars mark significant differences (p<0.01 and p<0.05, respectively, Student´s t-test) between ACC and NHS samples of the same time point. Error bars indicate SE.
A hit-and-run heat shock factor governs sustained histone methylation and transcriptional stress memory (2015)
Figure 2

Figure 2: Sustained induction of HS memory-related gene expression after an acclimatizing HS depends on HSFA2.

A. Transcript levels of the indicated HS-inducible genes after ACC in Col-0 (blue bars) and hsfa2 (orange bars) as determined by quantitative Reverse Transcription-PCR (qRT-PCR). Transcript levels were normalized to TUB6 and the respective NHS harvested at the same time point ([GENE OF INTERESTACC x h/ TUB6ACC x h]/ [GENE OF INTERESTNHS x h/ TUB6NHS x h]). Note that the y-axis is on a log10 scale. Error bars show SE of at least three biological replicates. Asterisks show significant differences between the genotypes at the same time point based on Student´s t-test (*: p<0.05; **: p<0.01).

B. Immunoblotting of HSP21 and HSP101 in Col-0 (C) and hsfa2 (h) at the indicated time points after ACC or NHS. TUBULIN was used as a loading control. M, marker.
A hit-and-run heat shock factor governs sustained histone methylation and transcriptional stress memory (2015)
Figure 3

Figure 3: Sustained H3K4me3 and H3K4me2 at HS memory-related loci depends on HSFA2.

H3K4me3 (A) or H3K4me2 (B) levels after an acclimatizing HS (ACC) in Col-0 and hsfa2 at HSP18.2, HSP22.0, HSP70 and APX2 as detected by ChIP-qPCR. Col-0 (blue bars) and hsfa2 (orange bars) seedlings were subjected to ACC or no treatment (NHS) 4 d after germination. At the indicated time points after the treatment, ChIP-qPCR was performed with antibodies against H3K4me3 (A), H3K4me2 (B) and H3 (for normalization). Schematics show positions of regions analyzed (grey bars, UTR; black bar, exons). Intergenic control region 1 is 3123 bp (APX2) or 2570 bp (HSP22.0) upstream of the TSS, or 5311 bp (HSP18.2) or 6725 bp (HSP70) downstream of the TSS, respectively. Data are averages over four biological replicates. Amplification values were normalized to input and H3 and the Col-0 4h NHS region 2 (HSP18.2, HSP22.0 and HSP70) or region 3 (for APX2). *, p<0.05; **, p<0.01 for differences between genotypes at the same time point and treatment; squares and triangles within bars mark significant differences (p<0.01 and p<0.05, respectively) between ACC and NHS samples of the same time point and genotype, Student´s t-test. Error bars indicate SE.
A hit-and-run heat shock factor governs sustained histone methylation and transcriptional stress memory (2015)
Figure 4

Figure 4: Histone H3K9ac profiles after ACC in Col-0 and hsfa2 at HSP18.2, HSP22.0, HSP70 and APX2 as detected by ChIP-qPCR.

Col-0 (blue bars) and hsfa2 (orange bars) seedlings were subjected to ACC or control treatment (NHS) 4 d after germination. At the indicated time points after the treatments, ChIP-qPCR was performed with antibodies against H3K9ac or H3. Schematics (grey bars, UTR; black bar, exons) show positions of regions analyzed. Intergenic control region 1 is 3123 bp (APX2) or 2570 bp (HSP22.0) upstream of the TSS, or 5311 bp (HSP18.2) or 6725 bp (HSP70) downstream of the TSS, respectively. Data are averages over four biological replicates. Amplification values were normalized to input and H3 and the Col-0 4h NHS region 2 (HSP18.2, HSP22.0 and HSP70) or 3 (for APX2). *, p<0.05; **, p<0.01 for differences between genotypes at the same time point and treatment; squares and triangles within bars mark significant differences (p<0.01 and p<0.05, respectively) between ACC and NHS samples of the same time point and genotype, Student´s t-test. Error bars indicate SE.
A hit-and-run heat shock factor governs sustained histone methylation and transcriptional stress memory (2015)
Figure 5

Figure 5: HSFA2 associates with HS memory-related genes transiently after an acclimatizing HS.

Binding of HSFA2-YFP to the promoter regions of HSP18.2 (A), HSP22.0 (B), APX2 (C), HSP70 (D) and ACTIN7 (E) was determined by ChIP-qPCR. Quantifications represent the enrichment relative to input in %. Red-orange-yellow bars represent time points (0.5 h-4 h-28 h) after a ACC and grey bars represent the respective NHS control samples. Schematics show the promoter region of the respective gene with regions analyzed (arrow, TSS; grey bar, 5’UTR; black bar, exon; black boxes, heat shock elements (5’-nnGnAnnTnCtn-3’). Regions are drawn to scale. Intergenic control region 1 is 4610 bp (HSP18.2) and 4192 bp (HSP70) downstream of the 3’UTR and 2570 bp (HSP22.0) or 3123 bp (APX2) upstream of the TSS, respectively. One representative experiment out of at least four biological replicates is shown. Error bars are SD of three replicates.
A hit-and-run heat shock factor governs sustained histone methylation and transcriptional stress memory (2015)
Figure 6

Figure 6: Expression profiles of HS memory-related genes after a recurring HS show HSFA2-dependent transcriptional memory

Response of selected HS-inducible loci upon recurring HS as determined by qRT-PCR. Transcript levels of APX2 (A), HSP18.2 (B), HSP22.0 (C), and HSP70 (D) in Col-0 (blue bars) and hsfa2 (orange bars) at the indicated time points after a single HS or two HS separated by 2 d. Plants were subjected to 37°C for 60 min on either d 4, d 6 or on d 4 + d 6 after germination. NHS samples were harvested at the same time points as HS d 4 samples. Transcript levels of four biological replicates normalized to TUB6 are shown. Small brackets compare the two genotypes at the same condition, large brackets compare the same genotype across two treatments (at 1 h after treatment) with Col-0 indicated first (Col-0/ hsfa2). Error bars show SE over four biological replicates. *, p<0.05; **, p<0.01; Student´s t-test.
Distinct biogenesis pathways may have led to functional divergence of the human and Drosophila Arglu1 sisRNA (2022)
Figure 1

Figure 1 - Characterization and biogenesis of Arglu1 sisRNA.

A Schematics showing the Arglu1 gene model and the different splicing isoforms.

B Top: Arglu1 gene locus indicating the primers' location (black arrow) and smFISH probes designed for Arglu1 sisRNA (green line) and Arglu1 pre-mRNA (magenta line). Bottom: RT-PCR showing the presence of Arglu1 sisRNA (P1+P2) detected from the different breast cancer cell lines.

C Experimental scheme for transcription inhibition treatment using alpha-amanitin and the indicated time points for cell harvesting.

D qPCR of Arglu1 sisRNA, pre-mRNA and mRNA expression levels normalized to gapdh from alpha-amanitin treatment at indicated time points. (n=3 biological replicates). Data are presented as mean ± SEM. **P≤0.01 (Student's t-test). See also Appendix Figure S1.

E RT-PCR indicating presence of Arglu1 sisRNA in the nuclear (N) and but not in the cytoplasmic (C) fraction. U85 gene was used as nuclear marker and 28S and 18S rRNA as cytoplasmic marker.

F smFISH of Arglu1 pre-mRNA (magenta) and Arglu1 sisRNA (green). Merged dots (white, arrowheads) represent Arglu1 pre-mRNA transcript. Scale bar: 20 μm.

G Quantification of Arglu1 pre-mRNA and Arglu1 sisRNA copy number from (F). (n = 50 cells) Cross, mean; Middle line, median; box, 25th to 75th percentiles; whiskers, minimum to maximum.

H Diagram indicating the locations of antisense morpholino oligonucleotides (AMOs) targeting the UCE region of Arglu1 intron 2 and primers used in (I).

I RT-PCR of Arglu1 sisRNA, spliced mRNA levels and actin as control after AMOs treatment.
Distinct biogenesis pathways may have led to functional divergence of the human and Drosophila Arglu1 sisRNA (2022)
Figure 2

Figure 2 - Arglu1 sisRNA is involved in autoregulation of its host gene in a UCE-dependent manner.

A Arglu1 gene locus indicating the locations of antisense oligonucleotides (ASOs) (red lines) designed for Arglu1 sisRNA KD and primers used in (B-D).

B, C qPCR of Arglu1 sisRNA and Arglu1 pre-mRNA expression levels in Arglu1 sisRNA ASO1 and ASO2 cells vs control cells. (n = 3 biological replicates).

D Chart showing the splicing indices (spliced/unspliced ratio) in Arglu1 sisRNA ASO1 and ASO2 cells vs control cells as measured by qPCR. (n = 3 biological replicates).

E Arglu1 gene locus indicating the location of antisense oligonucleotide (ASO) (red line) designed for Arglu1 sisRNA KD and primers used in (F).

F qPCR of Arglu1 mRNA expression levels at different exon junctions in Arglu1 sisRNA KD cells vs control cells. (n = 3 biological replicates).

G Left: Western blot analysis of ARGLU1 protein and GAPDH as loading control in control cells and Arglu1 sisRNA KD cells. Right: Quantification of relative ARGLU1 protein levels normalized to GADPH in Arglu1 sisRNA KD cells vs control cells from western blot analysis (left). (n = 3 biological replicates).

H Top: Schematic showing minigene expressing Arglu1 intron 2 wild type (WT) or intron 2 with UCE deletion (Δ) (location of UCE as indicated by orange bar). Bottom: Arglu1 gene locus indicating the primers' location (black arrows) for (I and J).

I Chart showing the splicing indices (spliced/unspliced ratio) in Arglu1 intron 2 WT OE cells and Arglu1 Intron 2 ΔUCE OE cells as measured by qPCR. (n = 3 biological replicates).

J qPCR of Arglu1 sisRNA levels in Arglu1 intron 2 WT OE cells and Arglu1 Intron 2 ΔUCE OE cells vs control cells (n = 3 biological replicates).

K, L qPCR of c-Myc and pS2 expression levels in 3 days Arglu1 sisRNA KD cells vs control cells under 100 nM estrogen treatment at indicated time points. (n = 3 biological replicates).

M Left: RT-PCR of AXIN1a and AXIN1b spliced levels and actin as loading control in 3 days Arglu1 sisRNA KD cells vs control cells under 100 nM estrogen treatment for 2 hrs. Right: Quantification of relative spliced AXIN1a/AXIN1b expression ratio normalized to actin from RT-PCR analysis (left). (n = 3 biological replicates).

N Schematics of Arglu1 gene autoregulatory loop model indicating involvement of Arglu1 sisRNA in promoting splicing of host gene (red arrow).

Data information: In (B-D, F, G, I-M), data are presented as mean ± SEM. ns=not significant, *P≤0.05, **P≤0.01 and ***P≤0.001 (Student's t-test). See also Fig EV1 and EV2.
Distinct biogenesis pathways may have led to functional divergence of the human and Drosophila Arglu1 sisRNA (2022)
Figure 3

Figure 3 - Arglu1 sisRNA binds ARGLU1 protein and inhibits its interaction with Arglu1 pre-mRNA.

A Top: Arglu1 gene locus indicating the primers' location (black arrows) for (A-C). Bottom: RIP-qPCR of Arglu1 pre-mRNA (P3+P4) and Arglu1 sisRNA (P1+P2) levels from ARGLU1 protein immunoprecipitation. Samples with beads and no antibodies were used as control. (n = 3 biological replicates).

B Left: Western blot analysis of ARGLU1 protein and GAPDH as loading control in 1 day Arglu1 sisRNA KD cells vs control cells. Right: Quantification of relative ARGLU1 protein expression level normalized to GADPH in 1 day Arglu1 sisRNA KD cells vs control cells from western blot analysis (left). (n = 3 biological replicates).

C RIP-qPCR of Arglu1 pre-mRNA and Arglu1 sisRNA levels normalized to gapdh in 1 day Arglu1 sisRNA KD cells vs control cells from ARGLU1 protein immunoprecipitation. (n = 3 biological replicates).

D Left: smFISH of Arglu1 sisRNA (green) coupled with immunostaining of ARGLU1 protein (red) shown in z-stack. Right: Zoom in z-slice on the boxed area showing co-localization (arrowheads) of Arglu1 sisRNA with ARGLU1 protein. The intensity plot shows the signal quantification and co-localization of Arglu1 sisRNA and ARGLU1 protein in the direction of the white arrow. Scale bar: 20 μm.

E Left: smFISH of Arglu1 pre-mRNA (green) coupled with ARGLU1 protein (red) immunostaining shown in z-stack. Right: Zoom in z-slice on the boxed area showing co-localization (arrowhead) of Arglu1 pre-mRNA with ARGLU1 protein. The intensity plot shows the signal quantification and co-localization of Arglu1 pre-mRNA and ARGLU1 protein in the direction of the white arrow.

F smFISH of Arglu1 sisRNA (green) and Arglu1 pre-mRNA (magenta) in 1 day Arglu1 sisRNA KD cells vs control cells. Scale bar: 20 μm.

G, H Quantification of Arglu1 sisRNA and Arglu1 pre-mRNA copy number from (F). (n = 90 cells). Cross, mean; Middle line, median; box, 25th to 75th percentiles; whiskers, minimum to maximum.

I Quantification of Arglu1 pre-mRNA and ARGLU1 protein co-localization from Arglu1 pre-mRNA smFISH coupled with ARGLU1 immunostaining in 1 day Arglu1 sisRNA KD cells vs control cells. (n = 5 biological replicates).

Data information: In (A-C, and G-I), data are presented as mean ± SEM. ns=not significant, *P≤0.05 and ***P≤0.001 (Student's t-test). See also Appendix Figure S2.
Distinct biogenesis pathways may have led to functional divergence of the human and Drosophila Arglu1 sisRNA (2022)
Figure 4

Figure 4 - Arglu1 sisRNA promotes robust ARGLU1 protein phase separation and localization to nuclear speckles.

A Immunostaining of MED1 (magenta) and SC35 (red) with transiently expressed mCherry-ARGLU1 protein (green) in MCF-7 cells. Zoom in of boxed area and the intensity plot shows the co-localization and signal quantification of mCherry-ARGLU1 protein, MED1 and SC35 in the direction of the white arrow. Scale bar: 20 μm.

B Confocal images of GFP-SRSF2 (green, control) and mCherry-ARGLU1 protein (red) at the pre-bleach, bleach, and post-bleach time points in FRAP analysis. The boxed area showing the speckle that was being bleached. Scale bar: 5 μm.

C Fluorescence intensities of the nuclear speckle over time in GFP-SRSF2 (green, control) and mCherry-ARGLU1 protein (red) FRAP analysis. (n = 3 separate nuclear speckles).

D Immunostaining of ARGLU1 protein (green) and SC35 (red) in 1 day Arglu1 sisRNA KD cells vs control cells. Zoom in of boxed area and the intensity plot shows the co-localization and signal quantification of ARGLU1 protein and SC35 in the direction of the white arrow. Scale bar: 20 μm.

E Confocal images of 125 nM ARGLU1-GFP protein (green) only and ARGLU1-GFP protein added with 50 nM IVT RNA (magenta), of either Arglu1 intron 2 with UCE (Intron 2 WT) or IVT Arglu1 intron without UCE (Intron 2 ΔUCE). White arrowhead indicates ARGLU1-GFP droplet incorporated with IVT Intron 2 WT and blue arrowhead indicates free ARGLU1-GFP droplet not incorporated with IVT Intron 2 WT. Scale bar: 10 μm.

F Quantification of total ARGLU1-GFP droplets from (E). (n = 3 biological replicates). Cross, mean; Middle line, median; box, 25th to 75th percentiles; whiskers, minimum to maximum.

G Quantification of average ARGLU1-GFP droplets size with/without incorporation of IVT intron 2 WT from (E, ARGLU1-GFP + Intron 2 WT). (n = 3 biological replicates). Cross, mean; Middle line, median; box, 25th to 75th percentiles; whiskers, minimum to maximum.

Data information: In (C), data are presented as mean ± SEM. ns=not significant and *P≤0.05 (Student's t-test). See also Figure EV3 and Appendix Figure S3.
Distinct biogenesis pathways may have led to functional divergence of the human and Drosophila Arglu1 sisRNA (2022)
Figure 6

Figure 6. dArglu1 sisRNA interacts with U1 snNRP, Arglu1 pre-mRNA and dARGLU1 protein in forming nuclear bodies and promotes dARGLU1 protein phase separation.

A Western blot analysis of dARGLU1 protein level from dARGLU1 protein immunoprecipitation in S2 cells. Samples with rabbit-IgG antibodies were used as control. ARGLU1 antibody #1, but not #2, worked for IP.

B RIP-qPCR of dArglu1 sisRNA, dArglu1 pre-mRNA, U1 snRNA and U85 snoRNA levels from dARGLU1 protein immunoprecipitation using ovary lysates. Samples with rabbit-IgG antibodies were used as control. (n = 3 biological replicates).

C-E RIP-qPCR of dArglu1 sisRNA, pre-mRNA and U1 snRNA levels normalized to U85 from dARGLU1 protein immunoprecipitation using dArglu1 sisRNA KD 1 and 2 ovaries vs control (sibling control, MTD-GAL4/CyO) ovaries. (n = 3 biological replicates).

F Left: Western blot analysis of dARGLU1 protein levels and ACTIN5C as loading control in dArglu1 sisRNA KD ovaries vs control (sibling control, MTD-GAL4/CyO) ovaries. Right: Quantification of relative dARGLU1 protein levels in dArglu1 sisRNA KD ovaries vs control ovaries from the western blot analysis. (n = 3 biological replicates)

G Left: Immunostaining of dARGLU1 protein (green) and VASA (red) in dArglu1 sisRNA KD germaria vs control (sibling control, MTD-GAL4/TM3) germaria. The asterisk (*) marks the anterior of the germarium. Right: Zoom in of the boxed area and the intensity plot shows the signal quantification of dARGLU1 protein and VASA in the direction of the white arrow. Scale bar: 10 μm.

H Quantification of germaria with and without nuclear speckle formation from dARGLU1 protein staining in dArglu1 sisRNA KD germaria vs control (sibling control, MTD-GAL4/TM3) germaria from (G). (n = 20 germaria)

I Confocal images of 125 nM dARGLU1-GFP protein (green) only and dARGLU1-GFP protein added with 50 nM IVT RNA (magenta), of either dArglu1 intron 2 until the 3' end of sisRNA (dArglu1 intron 2) or IVT dArglu1 intron 2 starting after 3' end of sisRNA (dArglu1 pre-mRNA). White arrowheads indicate dARGLU1-GFP droplets incorporated with IVT dArglu1 intron 2 and blue arrowheads indicate free dARGLU1-GFP droplets not incorporated with IVT dArglu1 intron 2. Scale bar: 10 μm.

J Quantification of total dARGLU1-GFP droplets from (I). (n = 3 biological replicates). Cross, mean; Middle line, median; box, 25th to 75th percentiles; whiskers, minimum to maximum.

K Quantification of average dARGLU1-GFP droplets size with/without incorporation of IVT dArglu1 intron 2 from (I, dARGLU1-GFP + dArglu1 intron 2). (n = 3 biological replicates). Cross, mean; Middle line, median; box, 25th to 75th percentiles; whiskers, minimum to maximum.

Data information: In (B-F), data are presented as mean ± SEM. ns=not significant, *P≤0.05, **P≤0.01 and ***P≤0.001 (Student's t-test). See also Figure EV5.