Fig. 3.

Pum1; Pum2 double knockout (DKO) ESCs have decreased self-renewal capacity. (A and B) Alkaline phosphatase (AP) staining of ESCs cultured under self-renewal and differentiation conditions as viewed by low (A) and high (B) magnifications. (C–F) Quantification of AP staining in different culture conditions. Percentage of cell-colony numbers are shown as mean ± SD. t-test: *P < 0.05, **P < 0.01. (G and H) Western blot analysis of pluripotency markers in wild-type, Pum1^−/−, Pum2^−/−, and Pum1^−/−; Pum2^−/− ESCs cultured under self-renewal (G) and differentiation (H) conditions. Boxes highlight lower protein levels of Nanog (blue), Sox2 (red), and Oct4 (green) in Pum1^−/−; Pum2^−/− ESCs compared to wild type, lower levels of Nanog (blue) in Pum1^−/− cells compared to wild type, and higher levels of all three pluripotency markers in Pum1^−/− ESCs compared to wild type.