Fig. 5: FBX8 targets HIF-1α, CDK4, and C-Myc for ubiquitin-mediated degradation.
From: FBX8 promotes metastatic dormancy of colorectal cancer in liver
a Western blot analysis was performed for SOX-9, SOX-2, Caspase-3, HIF-1α, C-myc, and CDK4 in subcutaneous tumors of SW480/MOCK, SW480/FBX8, SW620/NC, and SW620/shFBX8 cells, normalized to tubulin. b, c The half-life of HIF-1α, CDK4, and C-Myc were detected by a cycloheximide chase assay. d The expression of GST in E coli. detected by western blot. e–k E coli. transfected with an FBX8-expressing construct and GST-tagged IP of FBX8-Sec7 and FBX8-1 identified HA-HIF-1α-2, CDK4, and HA-C-Myc-2 as a binding domain. l SW620/NC, SW620/shFBX8, SW480/MOCK, SW480/FBX8, and SW480/FBX8 cells with MG132 treatment were co-transfected with plasmids expressing HA-Ubiquitin, and IP of HIF-1α, CDK4, and C-Myc followed by immunoblot analysis of HA-ubiquitin.
