Figure 3.

CtIP Depletion Impairs Human Cell Reprogramming

(A) Percentage of cells in sub-G1 phase in HFF cells or those reprogrammed to iPSCs bearing either the control shRNA shNT (white bars) or shCtIP (black bars). Error bars indicate ±SEM of three independent experiments. p Values were calculated using the t test. ns, not significant.

(B) Representative FACS plots of cell-cycle analysis in reprogrammed HFF cells of each condition. At least three independent experiments were performed.

(C) Reprogrammed HFF cells bearing the indicated shRNAs at day 10 were immunoblotted to analyze for the proactive and active forms of caspase-3. A representative western blot is shown of three independent experiments.

(D) Percentage of positive cells for γH2AX. Other details as in (A).

(E) Reprogramming efficiency of human cells containing an IPTG-inducible shRNA against CtIP (shCtIP-IPTG) or the control shNT-IPTG was analyzed in three independent experiments. Error bars indicate ±SEM. Statistical significance was performed using a t test.

(F) Representative images of hiPSC morphology in cells harboring the inducible shCtIP-IPTG obtained in the presence or absence of IPTG, as indicated.

(G) Western blot analysis against the CtIP and OCT4 proteins in HFF cells and hiPSC clones bearing the indicated shRNAs and in the presence (+) or absence (−) of IPTG. A representative western blot is shown of three independent experiments.

(H) Analysis of chromosomes per metaphase in iPSCs harboring IPTG-inducible shCtIP or shNT and reprogrammed in the presence (+) or absence (−) of IPTG. Tukey's multiple comparison test was used with n > 75.

See also Figure S2.