Fig. 6

FAM289-mediated TMZ resistance is through stem-like property acquisition via ERK pathway. (A) The apoptosis and cell cycle of U251 and U87-MG cells treaded with TMZ were analyzed by flow cytometry. The cell apoptosis was determined by Annexin V-fluorescein isothiocyanate (FITC)/PI staining. The percentages of Annexin-V-positive cells were indicated. (B) The cell cycle was analyzed by flow cytometry after PI staining. The data were processed with ModFit LT program. (C) The in vitro colony formation in cultured U251 and U87-MG cells. Images were taken on day 10 under an inverted microscope. (D) The cell lysates from FAM289-OE U251 and FAM289-KD U87 cells were immunoblotted with anti-stem-like property factors (OCT4, c-Myc, SOX2, CD133), anti-ERK, anti-pERK and anti-DNMTs antibodies. Corresponding wild type cells were used as controls. (E) The proportion of stemness factor CD133, Ki67 and Chemokine CXCR4 expression was analyzed by flow cytometry