Figure 1.

CME Is Required for the Maintenance of mESC Pluripotency and Self-Renewal

(A) Dot plot showing the results of the endocytic siRNA screen in mESCs, based on AP staining. Knockdowns resulting in a significant decrease in AP staining are marked in blue, while those resulting in an increase in AP staining are shown in red.

(B) Bright-field images show AP staining and morphology of mESCs 2 days post Cltc knockdown, or expression of K44A Dnm (K44A). Scale bar, 50 μm. NTi, non-targeting siRNA control; Cltci, Cltc siRNA; control, vector control.

(C) Bar graph showing mESC colony number, 3 days post indicated conditions.

(D and E) Line graph showing the proliferation rate of (D) mESCs and (E) MEFs over 3 days post indicated conditions.

(F) Cell-cycle analysis of mESCs and MEFs 3 days post indicated conditions. Bar graph shows the percent of cells in G1, S, and G2 phases of the cell cycle for both mESCs and MEFs.

(G) Bar graph showing the expression of pluripotency markers in mESCs under indicated conditions relative to control (n = 3). Control is shown as a dotted line at 1.

(H) Bar graph showing the expression of differentiation markers in embryoid bodies generated from Cltc knockdown mESCs by qRT-PCR analysis (n = 3). SCi, scrambled shRNA control; Cltci, Cltc shRNA.

Error bars represent mean ± SD from three independent experiments (n = 3). ^∗p < 0.05; ^∗∗p < 0.01; ^∗∗∗p < 0.001 by two-tailed Student’s t test.