Fig. 6: VAL abrogates Trim16-induced degradation of Vimentin.

a Co-IP assay following immunoprecipitation of Flag-tagged Vimentin shows the interaction of Vimentin with Trim16. b WB analysis of Vimentin expression in vector-control or Trim16-overexpressing A549 cells treated with or without MG132. c The effect of overexpressing Trim16 on the levels of K48-linked polyubiquitination of Vimentin was evaluated by immunoprecipitation of FLAG-tagged Vimentin in myr-AKT1-overexpressing A549 cells. d The half-life of Vimentin proteins were evaluated in vector-control or Trim16-overexpressing A549 cells treated with CHX and collected at the indicated time points. e Interaction between Trim16 and different truncated mutants of Flag-tagged Vimentin was evaluated by co-IP assays in 293FT cells. f The effect of overexpressing VAL in a dose-dependent manner on the interaction between Trim16 and Flag-tagged Vimentin as evaluated by immunoprecipitation of Flag. g The effect of overexpressing Trim16 on the levels of K48-linked polyubiquitination of Vimentin in the presence of overexpressed VAL or of ΔVAL was evaluated by immunoprecipitation of FLAG-tagged Vimentin in A549 cells. h WB analysis of Vimentin levels in A549 cells with the indicated treatments. i The effect of silencing VAL or together with silencing Trim16 on the levels of K48-linked polyubiquitination of Vimentin in A549-myr-AKT1 or LAD3 cells. j WB analysis of Vimentin levels in A549-myr-AKT1 or LAD3 cells silenced with VAL or together with Trim16 knockdown. Representative images derived from one of three independent experiments are presented. Relative protein levels were quantified by scanning densitometry and calculated as Band Intensity of Protein of Interest/Band Intensity of GAPDH (b, d). Source data are provided as a Source Data file.