Figure 3.

miR-302/367 Cluster Dually Regulates Cell Cycle and Apoptosis of hESC in a Dose-Dependent Manner

(A) Diagram of the gating strategy for hESC populations with different expression of TALE1-KRAB. hESCs were gated based GFP expression level (negative, low, medium, and high) and the following corresponding four populations were gated: P-I, GFP^negative; P-II, GFP^low; P-III, GFP^medium; P-IV, GFP^high. Low, Med, and High were hESCs with low GFP fluorescence, medium GFP fluorescence, and high GFP fluorescence, respectively.

(B) qPCR analysis of pri-miR-302/367 in four populations of hESCs expressing different level of TALE1-KRAB indicated by GFP fluorescence. Data are represented as mean ± SD of technical replicates (n = 3).

(C) A representative flow cytometry analysis of cell cycle and apoptosis in the four populations of cells with different degree of GFP fluorescence (P-I, P-II, P-III, P-IV).

(D) Cell cycle phase distribution of hESCs expressing different level of TALE1-KRAB. The four cell populations (P-I, P-II, P-III, P-IV) were gated as shown in (A) and analyzed as shown in (C), and their cell cycle distribution was calculated and shown as a percentage. Data are represented as mean ± SD of three independent experiments (^∗∗p < 0.01).

(E) Percentage of apoptotic cells in hESCs with a decreased expression level of miR-302/367 cluster. The apoptotic population of cells was represented by the sub-G0/G1 fraction of events in the four populations (P-I, P-II, P-III, P-IV). Data are represented as mean ± SD of three independent experiments (^∗∗p < 0.01).