Figure 3

Baf60b mediates 3TF-induced ATM activation. (A) The hepatic conversion was induced in wild-type (WT) TTFs transfected with shRNAs against the indicated genes. Forty-eight hours after 3TF transduction, p-ATM levels were imaged by the Operetta system and quantified by Harmony software. n = 2 independent experiments, and 350-500 nuclei were analyzed for each shRNA in both experiments. (B) Numbers of iHep colonies were quantified 8 days after 3TF transduction. Data represent two independent experiments. (C-F) The role of Baf60b in ATM activation and iHep formation was confirmed by shRNAs targeting Baf60b CDS (sh1-Baf60b) and 3′UTR (sh2-Baf60b). (C, D) p-ATM and p-p53 (Ser15) levels were analyzed by the western blot assay. (E) Cell proliferation was determined by BrdU incorporation and staining. n = 5 fields for each group. (F) iHep colony numbers were quantified 8 days after 3TF transduction. n = 4 independent experiments for each group. Data represent two independent experiments. Error bars indicate SD. ^*P< 0.05. Student's t-test.