Fig. 1 | Oncogene

Fig. 1

From: Mutant p53 gain of function underlies high expression levels of colorectal cancer stem cells markers

Fig. 1

Mutant p53 gain of function endows colorectal cancer cell lines with higher oncogenic potential. The colorectal adenocarcinoma-derived cell line, SW480 that endogenously expresses mutant p53R273H,P309S was stably infected with shRNA against p53 (shp53) or with shRNA against nonspecific sequence (shCon), as a control. a Western Blot displays mutant p53 protein levels. GAPDH serves as loading control. This figure combines two independent detections of p53 and GAPDH of the same gel. The two original images are presented in Supplementary Figure 1. b The shCon and shp53 SW480 cell lines (2 × 105 or 1 × 104) were injected into left and right limb of nude mice, respectively, followed by tumor growth surveillance. After 8–10 weeks mice were sacrificed and tumors size was measured. Asterisks denotes statistical significance (p < 0.01, n = 6). c Three independent newly infected shCon and shp53 SW480 cell lines were mixed and were injected (7 × 105) into left and right limb of nude mice, respectively, followed by tumor growth surveillance. After 5 weeks mice were sacrificed and tumors weight was measured. Asterisks denotes statistical significance (p < 0.05, n = 5). d Photos of tumors presented in c that illustrate the significant differences in the size of tumors obtained from injection of shCon and shp53 SW480 cells. e The established SW480 cell lines were treated with 5-FU (50 µM) for 72 h, followed by AnnexinV staining and ImageStream X analysis (Materials and methods). Graph in left panel indicates on percentage of apoptotic cells upon 5-FU treatment, normalized to NT cells. Right panel is a representative photo of an apoptotic cell. f RKO isogenic cell lines that express either WTp53 (RKO+/+) or mutant p53R248W (RKO+/m), or knocked-out for p53 (RKO−/−) [20] were treated with cisplatin (2.5 µg/ml) for 72 h, followed by AnnexinV staining and ImageStream X analysis. Graph in left panel indicates on percentage of apoptotic cells upon cisplatin treatment, normalized to NT cells. Right panel is a representative photo of an apoptotic cell. Graphs represent an average of three experiments. Error bars represent SE. Asterisk denotes statistical significance. For more representative photos obtained by ImageStream X see Supplementary Figure 13 A–B

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