Fig. 3: SE-enriched TFs rescue condensate localization in TRIM28-degraded mESCs.
From: Hijacking of transcriptional condensates by endogenous retroviruses
a, Genotype of the iPSC line and scheme of the experimental setup. The iPSC line contains degradation-sensitive Trim28-FKBP alleles and doxycycline-inducible Oct4, Sox2, Klf4 and c-Myc (OSKM) transgenes. b, Western blot validation of the FKBP degron tag and OSKM ectopic expression in iPSCs. c, Representative images of IAP RNA-FISH staining. The number and percentage refer to cells with detectable IAP foci, pooled from two biological replicates. Scale bars, 10 μm; inset scale bars, 2 μm. d. Quantification of cells with detectable IAP foci (IAP+ cells) at the indicated treatment regimes. e, IAP RNA expression is reduced in TRIM28-degraded iPSCs that ectopically express OSKM factors. The line plot shows qRT–PCR data of IAP RNA levels normalized to 0 h of dTAG-13 treatment. Data are from three independent biological replicates (three wells on a tissue culture plate) and are presented as mean values ± s.d. The experiment was repeated three times, and data from one representative experiment are shown. P value is from two-tailed t-tests. ***P < 1 × 10−4. f, Colocalization between the nascent RNA of miR290-295 and RNAPII puncta in TRIM28-degraded iPSCs that ectopically express OSKM factors. Separate images of individual z-slices (same z) of the RNA-FISH and IF signal are shown along with an image of the merged channels. The nuclear periphery determined by DAPI staining is highlighted as a white contour (scale bars, 2.5 μm). Also shown are averaged signals of either RNA-FISH or RNAPII IF centered on the miR290-295 RNA FISH foci or randomly selected nuclear positions. r denotes a Spearman’s correlation coefficient (scale bars, 0.5 μm). g, Elevated levels of miR290-295 SE transcript and Pri-miR290-295 nascent transcript in TRIM28-degraded iPSCs that ectopically express OSKM factors. qRT–PCR data was normalized to the 0 h of dTAG-13 treatment. Data are from three independent biological replicates (three wells on a tissue culture plate) and are presented as mean values ± s.d. The experiment was repeated three times, and data from one representative experiment are shown. P values are from two-tailed t-tests. *P = 0.027, ***P = 1 × 10−4.
