Fig. 1
BIR domain is required for XIAP-mediated EGFR protein expression and anchorage-independent growth in bladder cancer cells. a, b The cell extracts obtained from stable transfectants, T24T(Nonsense), T24T(shXIAP/Vector), T24T(shXIAP/ΔRING), UMUC3(Nonsense), UMUC3(shXIAP/Vector), UMUC3(shXIAP/ΔRING), UMUC3(shXIAP/Vector), or UMUC3(shXIAP/ΔBIR), were subjected to Western blot for determination of expression of XIAP and EGFR. GAPDH was used as the protein loading control. c, d The indicated stable transfectants were used for determination of their anchorage-independent growth ability in soft agar assay. Colonies with more than 32 cells were scored and presented as colonies/104 cells. Results were presented as means ± SD from triplicates. The asterisk “*” indicates a significant decrease as compared with nonsense transfectant, while symbol “※” indicates a significant increase in comparison to scramble vector transfectant (p < 0.05). Error bars represent S.D. e The cell extracts from T24T(shXIAP/Vector), T24T(shXIAP/EGFR-GFP), UMUC3(shXIAP/Vector), and UMUC3(shXIAP/EGFR-GFP), were subjected to Western blot for determination of indicated protein expression. β-actin was used as the protein loading control. f, g The indicated stable transfectants were used for determination of their anchorage-independent growth ability in soft agar assay. Colonies with more than 32 cells were scored and presented as colonies/104 cells. Results were presented as means ± SD from three independent experiments. The asterisk “*” indicates a significant increase as compared with the scramble vector transfectant (p < 0.05). Error bars represent S.D.