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Fig. 4 | Cell & Bioscience

Fig. 4

From: Inhibition of sphingolipid metabolism in osteosarcoma protects against CD151-mediated tumorigenicity

Fig. 4

CD151 depletion enhances c-myc ubiquitination and subsequent degradation. a Gene set enrichment analysis (GSEA) based on the RNA-seq using Hallmarks gene sets between WT and CD151 KO HOS-MNNG cells. b Transcriptional activity of c-myc was measured using luciferase promoter-reporter assay in CD151 overexpression and vector cells. Data (mean ± SD) are presented as fold changes in normalized luciferase activity. c RT-qPCR analysis of c-myc expression in HOS-MNNG and ZOSM cells silenced for CD151 expression. d and e The c-myc expression was detected in HOS-MNNG and ZOSM cells after knockout (d) or overexpression (e) of CD151 by Western blot. f Time-course analysis of c-myc protein levels in CD151 depletion HOS-MNNG cells. c-myc band density relative to β-actin was quantified, and the ratio of c-myc protein/actin protein was artificially set as 1.0 for samples untreated with CHX to obtain half-time (T1/2) of c-myc. g c-myc ubiquitination was analyzed in CD151 depletion cells. HOS-MNNG cells were transfected with the indicated plasmids followed by treatment with MG132 for 6 h. Cell extracts were immunoprecipitated with an anti-HA antibody, and ubiquitination c-myc was detected by western blot. Comparisons were made using the two-tailed, unpaired Student’s t-test; *p < 0.05, **p < 0.01, ***p < 0.001

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