Fig. 4

TRIM38 recognizes glucose transporter type 1 (GLUT1) and triggers ubiquitin-dependent degradation. a Gene Set Enrichment Analysis (GSEA) exhibited the enriched crosstalk in TRIM38-high versus TIM38-low samples. b Purification of TRIM38 proteins was conducted to identify the potential interacting substrates. The potential candidates with high frequency of enriched peptides were listed on the right table (c). d Western blot of indicated proteins in WCLs and Co-IP samples of IgG or anti-TRIM38 antibody obtained from the cell extracts of T24 cells treated with 20 μM of MG132 for 8 h. e Western blots of indicated proteins in WCLs from EJ cells transfected with an increasing amount of Myc-TRIM38 plasmids. The quantification of relative mRNA levels of GLUT1 was shown below, where the cells in the control group were transfected with the vector plasmid. f Western blot of GLUT1 and TRIM38 proteins in WCLs from T24 cells transfected with indicated plasmids with DMSO, MG132 (20 μM) or with Bortezmib (20 nM) for 12 h. g RT-qPCR assessment of GLUT1 mRNA expression in T24 cells treated with DMSO, MG132 (20 μM) or with Bortezmib (20 nM) for 12 h. The mRNA level of GAPDH was used for normalization. Data are shown as means ± SD (N = 3). The cells in control groups were treated with DMSO. h Western blot of the proteins of ubiquitination assays from T24 cells transfected with the indicated plasmids and treated with 20 μM MG132 for 12 h. i Representative IHC results exhibiting the negative associations between TRIM38 and GLUT1 proteins. Scale bars = 20 μm