Fig. 6: DRAK1 is degraded through K48-linked polyubiquitination induced by CUL3/SPOP E3 ubiquitin ligase. | Cell Death & Disease

Fig. 6: DRAK1 is degraded through K48-linked polyubiquitination induced by CUL3/SPOP E3 ubiquitin ligase.

From: Degradation of DRAK1 by CUL3/SPOP E3 Ubiquitin ligase promotes tumor growth of paclitaxel-resistant cervical cancer cells

Fig. 6

A Immunoprecipitation assay showing endogenous DRAK1 ubiquitination in HeLa and HeLa/PTX cells upon MG132 (10μM) treatment for 6 h. B Immunoprecipitation assay showing the interaction of Myc-DRAK1 with HA-CUL3 and Flag-SPOP in 293 T cells. C Immunoprecipitation assay showing the endogenous interaction of DRAK1 with CUL3 and SPOP in HeLa and HeLa/PTX cells. D In vivo ubiquitination assay showing Myc-DRAK1 ubiquitination by cotransfection of HA-CUL3 and Flag-SPOP in 293 T cells with MG132 (10 μM) treatment for 6 h. E In vivo ubiquitination assay showing Myc-DRAK1 ubiquitination by cotransfection of HA-CUL3 and Flag-SPOP in 293 T cells using Ni-NTA pull down of His-Ubiquitin WT or His-Ubiquitin 7KR mutant (mutating all seven lysines to arginines) with MG132 (10μM) treatment for 6 h. F Immunoprecipitation assay showing endogenous DRAK1 ubiquitin chain linkages. HA-CUL3 ectopically transfected in HeLa/PTX and indicated samples were immunoprecipitated with anti-DRAK1 antibody, followed by immunoblotting with anti-K48 Ub (K48-linkage specific Ubiquitin) or anti-K63 Ub (K63-linkage specific Ubiquitin) antibodies upon MG132 (10 μM) treatment for 6 h.

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