Fig. 1: Gas41 is highly expressed in undifferentiated mESCs.

a Western blot analysis of Gas41 and the pluripotent factors Oct4, Sox2 and Nanog at the specified times during embryoid body (EB) formation. β-Actin serves as a loading control. b RT–PCR analysis of Gas41, Oct4, Sox2 and Nanog expression levels during EB formation. Data was normalized to Actb with EB day 0 set as 1. Error bars represent the SEM from n = 3 biologically independent experiments. **P < 0.05; ***P < 0.001. Two-tailed unpaired Student’s t-test. c Dotplot depicting Gas41 mRNA levels in induced pluripotent stem cells (iPSC) and their parental somatic cells. Data were extracted from the Gene Expression Omnibus database GDS3842⁴⁹. d Heatmap showing change in gene expression (determined by qRT–PCR performed in technical duplicate) of Tip60/p400 and SRCAP complexes components on indicate day of RA treatment. Shown are log2FC gene expression at indicated day of treatment compared to day 0 (undifferentiated mESC). e Western blot analysis protein expression of indicated Tip60/p400 and SRCAP complexes components in RA treated mESCs. Arrow head, p400. β-Actin serves as a loading control. f Western blot analysis of Gas41 in control (shNT) and Gas41 KD (shGas41) mouse J1 ESCs. g RT–PCR analysis of Gas41 gene expression in control and Gas41 KD mESCs. Error bars represent the SEM from n = 3 biologically independent experiments. ***P < 0.001 (compared with shNT, Two-tailed Student’s t-test). h Phase contrast images of control and Gas41 KD mESCs. Bar, 200 μm. i Alkaline phosphate (AP) staining of control and Gas41 KD J1 cells. Bar, 200 µm. j Stacked bar plot showing quantification of AP staining from i. Error bars represent the SEM from 6 (shGas41-1) or 7 (shNT and shGas41-2) randomly selected fields. Statistical analyses are available in Supplementary Table S7