Figure 2.

CtIP Deficiency Impairs Mouse Cell Reprogramming

(A) Doxycycline-inducible OSKM MEFs were transduced with shCtIP and shNT lentivirus prior to induction with doxycycline. Reprogramming efficiency was analyzed by counting the number of colonies in triplicate of each biological sample. Error bars indicate ±SEM of a biological triplicate.

(B) Representative pictures of miPSC morphology.

(C) The sub-G1 peak was quantified in MEFs (–Dox) or reprogrammed MEFs at day 10 (+Dox) by FACS analysis in cells downregulated for CtIP (black bars) or expressing a control shRNA (white bars). Error bars indicate ±SEM of three independent experiments.

(D) Representative cell-cycle plots of reprogrammed MEFs in each condition. At least three independent experiments were performed.

(E) Immunoblot of proactive and active/cleavage caspase-3 in reprogrammed MEFs at day 10 (+Dox) bearing the indicated shRNAs. A representative western blot is shown of three independent experiments.

(F) FACS quantification of cells positive for γH2ax. Other details as in (C).

(G) The indicated proteins were immunodetected in samples from OSKM-induced MEFs after 10 days of continuous doxycycline treatment. A representative western blot is shown of three independent experiments.

t Test statistical analyses in (A), (C), and (F) were performed using at least three independent experiments.

See also Figures S2 and S3.