Fig. 5

INPP5F activates Notch signaling pathway through interacting with ASPH. A The base-peak plot of mass spectrometry analysis of protein complex immunoprecipitated by anti-FLAG antibody in Huh7 overexpressing FLAG-INPP5F. The INPP5F pulled-down peptide of ASPH was indicated. B Lysates of Huh7 transiently overexpressing FLAG-INPP5F or HA-ASPH were immunoprecipitated for FLAG or HA and immunoblotted for ASPH or INPP5F, respectively. C Western blot was performed to investigate the influence of INPP5F on the expression of ASPH, NICD, HES1 and HEY1 in SK-Hep1, MHCC-97H and Huh7 cells. (D-G) Huh7 overexpressing INPP5F were transfected with ASPH siRNA for 24 h, and then were subjected to (D) QRT-PCR, (E) western blot, (F) EdU assay, and (G) cell cycle analysis. Knockdown of ASPH significantly attenuated INPP5F-enhanced expression of HES1, HEY1, cyclin E1 and c-MYC, as well as cell proliferation and G1/S phase transition. Data is presented as means ± standard error for three independent experiments, *P < 0.05, ns: not significant