Fig. 5: DRAK1 protein is degraded by CUL3/SPOP E3 ubiquitin ligase in paclitaxel-resistant cells. | Cell Death & Disease

Fig. 5: DRAK1 protein is degraded by CUL3/SPOP E3 ubiquitin ligase in paclitaxel-resistant cells.

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

Fig. 5

A Immunoblot analysis showing DRAK1 expression in HeLa and HeLa/PTX cells with or without MG132 (10μM) treatment for 6 h and β-actin was used for internal control. B List of DRAK1 binding protein candidates analyzed from the mass spectrometry-based crosslinking assay. C RT-PCR (left) and immunoblot analysis (right) showing CUL3 and DRAK1 expression in HeLa and HeLa/PTX cells. 18 s or β-actin was used for internal control, respectively. D Immunoblot analysis showing Myc-DRAK1 expression by cotransfection of HA-CUL3 and its adaptor proteins, Flag-KLHL20, Flag-KEAP1, and Flag-SPOP in 293 T cells. E Immunoblot analysis showing the expression of Myc-DRAK1 by dose-dependent changes in the expression levels of Flag-SPOP with HA-CUL3. F Immunoblot analysis showing Myc-DRAK1 expression by cotransfection of HA-CUL3 and Flag-SPOP with or without MG132 (10 μM) treatment for 4 h. G Immunoblot analysis showing endogenous DRAK1 expression in HeLa and HeLa/PTX cells transfected with CUL3-siRNAs and SPOP-siRNAs. H qRT-PCR showing the expressions of downregulated target genes by DRAK1 overexpression validated from Fig. 4C in samples from G.

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