Fig. 6.
Defective XCI in female Stc1−/− mice is not accompanied by increased X-linked gene expression. (A) Schematic of the RNA-Seq analysis pipeline. (B) Distribution of log2-transformed ratio of X-linked gene expression in MEFs from female Stc1−/− (KO) and Stc1+/+ (WT) embryos (n = 3 per genotype). (C) Box plot of X-linked gene expression (log2-transformed FPKM) in MEFs from female Stc1−/− and Stc1+/+ embryos (n = 3 per genotype). The boxed areas span the first to the third quartile. The whiskers represent 15th and 85th percentiles. (D) qRT-PCR analysis monitoring expression of Mecp2 and Hprt in MEFs from two different litters of female Stc1−/− and Stc1+/+ embryos (n = 2 mice total per genotype). The results were normalized to those obtained in Stc1+/+ MEFs, which was set to 1. Error bars indicate SE. (E) Immunoblot showing MECP2 and STC1 levels in female Stc1+/+ and Stc1−/− MEFs (Left) or brain tissue female Stc1+/+ and Stc1−/− P1 mice (Right) (n = 3 per genotype). α-Tubulin (TUBA) was monitored as a loading control. (F) qRT-PCR analysis of Stc1, Xist, Mecp2, and Hprt expression in BMSL2 cells expressing a NS or STC1 shRNA. The results were normalized to those obtained with the NS shRNA, which was set to 1. Error bars indicate SE. (G) Immunoblot showing MECP2 and STC1 levels in BMSL2 cells expressing a NS or Stc1 shRNA.