Fig 5

SVCV-G protein interacts with hif1αa and hif1αb to attenuate K48-linked polyubiquitination of hif1αa and hif1αb. (A) Myc-hif1αa co-localized with Flag-SVCV-G as revealed by co-localization assays. Scale bar = 10 µm. (B) Myc-hif1αb co-localized with Flag-SVCV-G as revealed by co-localization assays. Scale bar = 10 µm. (C) Myc-hif1αa interacted with Flag-SVCV-G as revealed by co-immunoprecipitation assays. (D) Myc-hif1αb interacted with Flag-SVCV-G as revealed by co-immunoprecipitation assays. (E) Endogenous interaction between hif1α and SVCV in ZFL cells. Anti-SVCV-G antibody was used for immunoprecipitation (IP), and the interaction was detected by IB using anti-hif1α antibody. (F, G) Ubiquitination analysis of hif1αa in HEK293T cells transfected with Myc-hif1αa, Flag-SVCV-G (Flag empty vector [-] was used as a control), and His-Ub (F) or His-Ub-K48 (G) for 24 h, and then treated with MG132 (20 µM) for 8 h. (H, I) Ubiquitination analysis of hif1αb in HEK293T cells transfected with Myc-hif1αb, Flag-SVCV-G (Flag empty vector [-] was used as a control), and His-Ub (H) or His-Ub-K48(I) for 24 h, and then treated with MG132 (20 µM) for 8 h.