Fig. 1

CHIP-silencing enhances AGS cell growth. a The cell growth curves of the sictrl and siCHIP cells were detected by an x-Celligence system. E-plates were plated with 4000 cells/well and cell growth was continuously monitored for 72 h. b The anchorage-independent growth of the sictrl and siCHIP cells were detected using soft agar. 24-well plates were plated with 10,000 cells/well. Colonies were counted after 14 days’ incubation. c The bar chart represents the frequencies of Ki-67⁺ cells of the two established cell lines. d Western blotting analysis of the protein expression of total ERK1/2 and p-ERK1/2 in siCHIP and sictrl cells. Actin was used as an internal control. e Cell viability assay. 96-well plate was plated with 4000 cells/well. OD570 was measured by means of spectrophotometer microplate reader at 24, 48 and 72 h. f A TUNEL assay was carried out to quantitatively analyze the apoptotic cells. The bar chart represents the percentages of apoptotic cells. g Western blotting analysis of apoptosis-associated proteins. Actin was used as an internal control. h Cell cycle assay was determined by flow cytometry analysis using PI-staining. The histograms represent the percentages of cells in G₀–G₁, S, and G₂-M phases. i Western blotting analysis of cell cycle-related proteins. Actin was used as an internal control