Fig. 4

c-Myc is ubiquitinated substrate of β-TrCP. A. The interaction between β-TrCP and c-Myc proteins was examined using immunoprecipitation expriments in CNE2 and 5-8F cells after transfected with Flag tagged c-Myc. B. Immunofluorescence experiments showed that β-TrCP and c-Myc were co-localized in CNE2 and 5-8F cells. blue: DAPI-stained nucleus; green: anti-β-TrCP; red: anti-c-Myc; yellow: co-localization of β-TrCP and c-Myc. The merged image represented the overlap of DAPI, β-TrCP, and c-Myc. Scale bar, 20 μm. C. The interaction between c-Myc and the WD40 domain of β-TrCP was examined in CNE2 and 5-8F cells after transfected with Flag-tagged full-length β-TrCP or truncated fragments (F-box or WD40 domain) by immunoprecipitation using anti-Flag antibody. D. The interaction between β-TrCP and phosphorylated c-Myc proteins was examined in CNE2 and 5-8F cells by immunoprecipitation with anti-β-TrCP or anti-phosphorylated (T58 + S62) c-Myc antibodies. E. Wound healing assay showed that overexpression of c-Myc reverses the migrative ability of β-TrCP in NPC cells. Data were represented as mean ± SD. ***, p < 0.001, ns, not significant. F. Transwell assay showed that overexpression of c-Myc reverses the invasive ability of β-TrCP in NPC cells. Data were represented as mean ± SD. ***, p < 0.001, ns, not significant