Fig. 4: VCP maintains FASN stability in OS. | Cell Death & Disease

Fig. 4: VCP maintains FASN stability in OS.

From: VCP enhances autophagy-related osteosarcoma progression by recruiting USP2 to inhibit ubiquitination and degradation of FASN

Fig. 4

A, B Western blot assay was utilized to monitor the changes in FASN protein levels, after overexpression (A) or knockdown (B) of VCP, respectively, treatment with CHX (50 μg/mL, 6 h) or MG132 (20 μM, 6 h) in143B and U2OS cells. C Representative images (left) and the quantification (right) of western blot was performed to evaluate the half-life of FASN protein in 143B and U2OS cells. All groups were treated with cycloheximide (CHX, 50 μg/mL), a classic protein synthesis inhibitor, from 0 – 18 h. D The total level of FASN ubiquitinated proteins in OS cells was detected by western blot, 143B and U2OS cells were treated with Ca-VCP or Ci-VCP. E Levels of exogenous FASN ubiquitination in 293 T cells co-transfected with MYC-tagged VCP, His-tagged Ub and HA-tagged FASN. F 293T cells were transiently transfected with plasmids encoding MYC-tagged VCP, HA-tagged FASN, and His-tagged ubiquitin mutants (K11, K48 or K63). FASN ubiquitination was then analyzed by WB.

Back to article page