Fig. 1

CHIP silencing inhibits cell growth of DLD-1 cells. a The cell growth curves of the siCHIP and sictrl cells were detected using a real-time x-Celligence system. E-plate was plated with 10,000 cells/well and the cell growth was continuous monitored for 72 h. Significant differences were indicated (Student’s t test, **P < 0.01, ***P < 0.001). b The apoptosis of the siCHIP and sictrl cells was determined by the AnnexinV/PI staining. AnnexinV positive and AnnexinV/PI positive cells were determined as apoptosis cells. Values ± SD from three individual experiments were displayed. c Cell proliferation of the siCHIP and sictrl cells was determined by the Brdu proliferation assay. 96-well plate was plated with 10,000 cells/well and was added 10 μl buffer after cultured for 24, 48 and 72 h. OD450 was measured using spectrophotometer microplate reader. Significant differences were indicated (Student’s t test,***P < 0.001). d CHIP depletion caused cell cycle arrest at the G0–G1 phase. Cell cycle parameters of siCHIP and sictrl cells were determined by flow cytometry analysis using propidium iodide. The histograms report the percentage of cells in G0/G1, S and G2/M phases of all the establishing cell lines. ***P < 0.001. e Western blotting analysis of the protein expression of total ERK1/2, p-ERK1/2, p38, p-p38, as well as total AKT andp-AKT⁴⁷³ and p-AKT³⁰⁸ in siCHIP and sictrl cells. Actin was used as an internal control. f Western blotting analysis of the protein expression of the NF-κB subunits. Actin was used as an internal control. g Western blotting analysis of the protein expression of the cell cycle-related protein cyclinD1. Actin was used as an internal control