Figure 2.

JPX RNA is dispensable for XIST expression in human. (A) JPX genomic sequence is weakly conserved, as illustrated by the conservation score in 100 vertebrates and alignment of the human genomic region to five other mammalian species. Red shade highlights the JPX locus. (B) JPX escapes XCI in female fetal fibroblasts and primed H9 hESCs, as assessed by JPX/XIST double RNA-FISH. (C) Scheme of hLGs lipofection strategy in primed H9 hESCs and of JPX RNA isoforms (red line: LG-targeted exon). (D) JPX RNA levels were reduced following hLGs transfection, RT-qPCR, n = 3. (E) XIST RNA levels were unaffected following JPX RNA depletion in primed H9 hESC, RT-qPCR, n = 3. (F-G) JPX KD did not affect the number of cells expressing XIST (chi-square test) nor the volume of XIST RNA territory, RNA-FISH, Mann-Whitney test. Scale bars are 5 μm. Unless stated otherwise, unpaired two-tailed t-test were performed and error bars represent standard deviation; n.s., not significant; *P < 0.05, **P < 0.01 and ***P < 0.001. Number of counted cells is in brackets.