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Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: The novel circSLC6A6/miR-1265/C2CD4A axis promotes colorectal cancer growth by suppressing p53 signaling pathway

Fig. 3

C2CD4A promoted CRC cells growth via p53 signaling pathway and interacted with p53. a The heatmap and scatter diagram showed the differential expressed mRNA between RKO/sh-C2CD4A (KD-1, 2, 3) and RKO/sh-Ctrl cells (NC-1, 2, 3) screened by human cDNA microarray. b Top 10 pathways of KEGG analysis based on DEGs. c IP/MS analysis of Flag-C2CD4A-associated proteins. The Flag peptide elution was resolved on SDS-PAGE and Coomassie blue staining. d Several proteins related to cell apoptosis identified from Flag-C2CD4A IP/MS analysis. e The relative mRNA expressions of C2CD4A, p53, CDKNIA(p21), BAX, FAS, SESN1 and TiGAR were detected using qRT-PCR. f The levels of C2CD4A, p53, CDKNIA(p21) and BAX protein expression were detected using western blot. g 293 T cells were co-transfected with Flag-C2CD4A and HA-P53 at various concentrations, and then Flag and HA expression were detected using western blot. h HCT116 cells were treated with Dox at 1, 2 μM for 24 h, and treated at 1 μM for 12, 24 h. C2CD4A and p53 expressions were detected using western blot. i The lysates of RKO cells were transfected with Flag-C2CD4A followed by immunoprecipitated using anti-IgG, anti-Flag or anti-p53 antibodies, and immunoprecipitation products were analyzed by western blot with indicated antibodies. l 293 T cells were co-transfected with Flag-C2CD4A and HA-p53. Immunoprecipitation was carried out using anti-Flag or anti-HA antibody, and immunocomplexes were detected using indicated antibodies. (ns showed no significance. *P < 0.05, **P < 0.01)

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