Fig. 4.

TC2N involves in regulating p53 signaling pathway. a Identification of potential pathways downstream of TC2N with the TCGA lung cancer RNAseq (IlluminaHiSeq; n = 1124) data set, as described in the Materials and methods. b Heatmaps for correlations between TC2N and p53 signaling pathway-related genes in the TCGA lung cancer RNAseq (IlluminaHiSeq; n = 1124) data set. Correlation coefficient R and P values were calculated by Spearman's correlation analysis. c qRT-PCR analysis of P53, P21, BAX and Bcl-2 expression in H460 cells transiently transfected with the negative control or TC2N shRNA. ACTIN serves as an internal control. d The protein levels of TC2N, p53, P21, BAX and Bcl-2 were monitored by WB after knockdown of TC2N in H460 cells. e qRT-PCR analysis of P21, BAX and Bcl-2 expression in H1299 cells transiently transfected with the negative control or TC2N shRNA. f The protein levels of TC2N, p53, P21, BAX and Bcl-2 were monitored by WB after knockdown of TC2N in H1299 cells. ACTIN serves as an internal control. g The effects of TC2N knockdown on the p53 response reporter construct pp53-TA-Luc in H460 cells. h The effects of TC2N overexpression on the p53 response reporter construct pp53-TA-Luc in A549 cells. i H1299 (p53 null) cells were co-transfected with p53 response reporter construct pp53-TA-Luc, negative control or TC2N shRNA and HA-p53 expression vectors. Luciferase activity was measured at 24 h after the transfection. j H460 cells were co-transfected with PG13-luc or MG15-luc reporters and TC2N expression vectors as indicated, and luciferase activity was measured 24 h after transfection. Results are means ± SEM of three independent experiments **P < 0.01