Fig. 7

DIP2A knockdown abolished the effects of Fstl1 on TMZ resistance in vitro and in vivo. A and B, D54 cells cotransfected with Fstl1 or vector, and shDIP2A were treated with TMZ (200 μM) for 48 h. Flow cytometry was used to measure cells apoptosis (a), western blot analysis of the indicated proteins expression (b). c D54/DIP2A-KD cells (D54 cell with DIP2A knockdown) transfected with Fstl1 or vector were treated with different doses of TMZ for 48 h or 200 μM TMZ at indicated times. Cell proliferation was evaluated using CCK8 assay. d Colony formation assays were done with D54 cells cotransfected with Fstl1 or vector, and shDIP2A in the presence of TMZ (200 μM). e Representative images of tumors originated from D54/DIP2A-KD cells stably expressing Fstl1 or vector control in the presence of TMZ (66 mg/kg/day by oral gavage) on the 42nd day. f The level of MGMT, γ-H2AX, and cleaved caspase-3 was examined by IHC analysis. g Growth curve of subcutaneous tumor xenografts. h Tumor weight is the means of three independent experiments±SEM. i Representative pseudocolor bioluminescence images of intracranial xenografts bearing P-GBM2/DIP2A-KD cells transfected with shFstl1–7 or shCtrl in the presence of TMZ on the days as indicated. j IHC analysis of MGMT, γ-H2AX, and cleaved caspase-3 expression in intracranial xenografts. k Survival curve of P-GBM2/DIP2A-KD cells-derived intracranial xenograft tumors treated with shFstl1–7 or shCtrl in the presence of TMZ (n = 6). Student’s t tests were performed. Data are presented as mean ± SEM (^#P > 0.05), scale bar = 100 μm