Fig. 4. CCT8 inhibits the entry of WTp53 into the nucleus and antagonizes its anti-tumor effect.

A GSEA demonstrated enrichment of p53 signal pathway in high CCT8 expression CRC group. (p = 0.012). B The Co-IP was performed to analyze the endogenous interaction between CCT8 and p53 in CRC cells. Cells were lysed and purified by anti-CCT8 or anti-p53 affinity gel; protein pellets were analyzed by Western Blot with anti-p53 and anti-CCT8 antibodies. C Effect of CCT8 on the expression of p53 were detected by Western Blotting analysis (a representative set of data was shown, n = 3). D Immunofluorescence staining showed that CCT8 could inhibit the entry of p53 into the nucleus. The arrow shows nuclear p53. The box areas highlighted the co-localization between CCT8 and p53. E Paraffin-embedded CRC sections were stained with anti-CCT8, anti-LASP1 or anti-p53 antibodies. Visualization of two representative cases was shown. F Paraffin-embedded CRC sections were stained with anti-MTp53 antibodies. Visualization of two representative cases was shown. G Graphical illustration of statistical LASP1, CCT8 and p53 distribution in CRC tissues. The high expression of p53 in cytoplasmic is more frequently accompanied with high CCT8 in WTp53 cases (p < 0.05). H Graphical illustration of statistical CCT8 and MTp53 distribution in CRC tissues. The expression CCT8 was in-dependent of MTp53 expression (NS). I Scatter plot display of the correlation between CCT8 and MTp53(NS). J Scatter plot display of the correlation between CCT8 and p53.