Fig. 3: VCP cooperates with FASN to facilitate its expression. | Cell Death & Disease

Fig. 3: VCP cooperates with FASN to facilitate its expression.

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

Fig. 3

A Overlapped results combining autophagy database and FASN mass spectrometry analyses. B Silver staining assay(left) to detect proteins cooperating with FASN proteins and VCP peaks in protein mass spectrometry(right). C, D Validation of VCP-FASN interaction in 143B, U2OS and 293 T cells using CoIP assay. E, F Representative images (left) and the quantification (right) of immunofluorescence demonstrate the localization of FASN and VCP proteins in 143B and U2OS cells. G VCP truncated structure. The wild-type VCP, consisting of 806 amino acids, is divided into four structural domains: ΔN, ΔD1, ΔD2, and ΔC tail. Gray areas indicate deleted regions. H 293 T cells were transfected with MYC-labeled full-length VCP and its mutants with deletions in each region. Cell lysates were immunoprecipitated with anti-MYC antibody. Western blot assay was conducted to identify the expression of the MYC probe and FASN. I Western blot assay was performed to identify alterations in FASN protein levels resulting from stable transfection of Ca-NC, Ca-VCP, Ci-NC, Ci-VCP#1, or Ci-VCP#2. J CO-IP assay was utilized to measure the potency of FASN interactions with VCP, following treatment with EBSS for durations of 0, 2, 4, and 6 h.

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