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. 2022 May 31;21:37–45. doi: 10.1016/j.reth.2022.05.005

Fig. 2.

Fig. 2

Pluripotent marker gene expression in hetKO iPSCs was comparable with isogenic control line. (A) The expression of undifferentiated marker genes, including POU5F1, NANOG, SOX2, DNMT3B, TDGF1, and TRIM28, was comparable between control isogenic line and Homo sapiens (hsa)-miR-302/367 cluster heterozygous knockout Edom_iPSCs. Primary microRNA (miR) of hsa-miR-302/367 (pri-miR-302) was reduced by half in heterozygous knockout Edom_iPSCs. (B) The expression of mature miRs from the hsa-miR-302/367 cluster was analyzed by a TaqMan microRNA assay system in heterozygous knockout Edom_iPSCs and isogenic control lines. Mature miR-302a/b/c/d/and miR-367 were reduced by half in heterozygous knockout Edom_iPSCs. (C) Undifferentiated markers, including POU5F1, NANOG, and stage-specific embryonic antigen 4 (SSEA4), were normally expressed and localized in heterozygous knockout Edom_iPSCs and isogenic control lines. 4′,6-diamino-2-phenylindole (DAPI) staining is shown in blue. Scale bars are 100 μm. (D) The cell proliferation rate was comparable between the control isogenic line and hsa-miR-302/367 cluster heterozygous knockout Edom_iPSCs. (E) Apoptosis and cell-cycle related marker gene expression was comparable between the control isogenic line and hsa-miR-302/367 cluster heterozygous knockout Edom_iPSCs. hetKO, hsa-miR-302/367 cluster heterozygous knockout; iPSC, induced pluripotent stem cell; wt, wild-type. ∗P < 0.05. ∗∗P < 0.01.