Figure 4 .

FBXO32 promoted ATR ubiquitination and degradation

(A) Overexpression and knockdown of the FBXO32 gene in MG63 cells were confirmed by western blot analysis. Results showed that FBXO32 knockdown significantly increased ATR expression and high expression of FBXO32 suppressed the expression of ATR. (B) Western blot analysis and qPCR results indicated that the level of FBXO32 in OS cells decreased at 12 h and increased at 24 h after irradiation. (C) Co-IP assay demonstrated the binding of FBXO32 with ATR (MG63 cells were transfected with ATR-HA and FBXO32-Flag plasmids; IgG served as a negative control). (D) MG63 cells were transfected with plasmids (shFBXO32, ATR-HA, FBXO32-Flag and Ub-His) for 48 h followed by treatment with 20 μM MG132 for 8 h. Western blot analysis showed that FBXO32 promoted ATR ubiquitination and degradation. ** P<0.01.