Fig. 1

POGZ depletion leads to loss of ESC self-renewal. A (Up) Cartoon depicting the gRNA targeting sites at exon 2 of the mouse Pogz gene; (Bottom) Genotyping showing the mutant allele 1 (Mut1) of Pogz gene. B qRT-PCR showing the reduction of Pogz expression in mutant ESCs. C Western blot analysis of POGZ in control and Pogz−/− ESCs. D Double IF staining of OCT4 and POGZ showing that OCT4 levels were not significantly altered in early passage mutant ESCs. E The cell growth cure showing the proliferation defects of Pogz−/− and Pogz+/− ESCs compared to control ESCs. Mut1 ESCs at passage 12 were used for the assay. F Representative image showing morphology of control and late passage Pogz−/− ESCs (passage number is 18). White arrows pointing to the flattened differentiating cells. G Alkaline phosphatase staining of control and Pogz−/− ESCs. White arrows pointing to cells with reduced AP activities. H The mRNA expression levels of representative pluripotency-related, mesodermal, neuroectodermal, endodermal genes in control and Pogz−/− ESCs. I IF staining of GATA6 showing that GATA6 was abnormally expressed in Mut1 Pogz−/− ESCs. Restoring POGZ rescued the abnormal expression of Gata6. J WB showing that OCT4 levels were reduced in Pogz−/− ESCs, which can be reversed by reintroducing FLAG-POGZ. Bar: 25 μm. K Volcano plot showing the up- and down-regulated genes in the control and early passage Pogz−/− ESCs. The RNA-seq experiments were repeated two times. L GO analysis of up-regulated DEGs. M GO analysis of down-regulated DEGs. All WB and IF were repeated at least two times. qRT-PCR was repeated three times