Fig. 6

a, Expression level of SPEN in naive (PXGL and NaiveCult) and primed H9 cells obtained from RNA-seq data (n=3). Data are presented as the mean values±s.d. of replicates; Wilcoxon P value≥0.05 (NS). b, RT–qPCR showing fold enrichment levels of XIST, β-actin and MALAT1 normalized to RPLP0 following RIP of SPEN (n=3). Data are presented as the mean values±s.d. of replicates; Wilcoxon P value<0.05 (*). c, RT–qPCR analysis of SPEN expression after siRNA KD using two different mixes of siRNAs (SPEN1 and SPEN2; n=3). Values are normalized to β-actin. Data are presented as the mean values±s.d. of replicates; Wilcoxon P value≥0.05 (NS). d, RT–qPCR analysis of XIST expression after SPEN KD (n=3). Values are normalized to β-actin. Data are presented as the mean values±s.d. of replicates; Wilcoxon P value≥0.05 (NS). e, Percentage of biallelically expressed SNPs from X chromosome in scramble (n=3) and SPEN (n=3) KD naive H9 cells, obtained from RNA-seq data. Box plots represent the median (center), first and third quartiles (hinges) and ±1.5 IQR (whiskers). f, pXi allelic ratio from RNA-seq data of scramble and SPEN KD naive H9 hES cells (n=3). Data are presented as the mean values±s.d. of replicates. g, Heatmap showing the pXi allelic ratio of XIST-sensitive and XIST-resistant gene expression upon SPEN KD in naive H9 hES cells. An increase in pXi allelic ratio (FC>1.2) from one (*) or both (**) siSPEN mixes is indicated. Unless otherwise specified, P values were calculated by a two-sided unpaired t-test. Levels of significance: NS (P≥0.05), *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001.

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