Fig. 5

The lysine 150 and 169 sites in exon 6 affect total CDC25A ubiquitination and subcellular localization. A Both K150 and K169 were responsible for CDC25A ubiquitination. Single or double ubiquitin site mutations (K150R, K169R, and K150/169R) of CDC25A(L) were conducted. The immunoprecipitation results revealed less ubiquitination levels in both single-mutated situations, and double mutation further enhanced this effect. The HIS level showed no change of other proteins in these mutation systems, and the GFP input showed more CDC25A(△E6) expression in single- or double-mutated conditions. B Subcellular localization of the CDC25A(L) WT and mutated plasmids. Significantly stronger nucleus signals were detected in single- and double-mutated conditions. Scale bars, 20 μm. C Mutations in K150 and K169 enhanced foreign CDC25A(L) protein stability. HCC cell lines with WT and mutated GFP-tagged CDC25A(L) transfection were introduced into the CHX experiment at indicated time points