Fig. 5
USP10 interacted with and stabilized HDAC7. a USP10 and HDAC7 interacted in NSCLC cells. b The statistical analysis of relative HDAC7 mRNA levels in USP10 overexpression and knockdown cells. mRNA was normalized to GAPDH. c Representative western blotting of USP10, HDAC7 and FGF18 in indicated cells. d Protein stability analysis showing a reduced half-life of HDAC7 in USP10 knockdown cells. e Knockdown of USP10 promoted ubiquitination of HDAC7 in NSCLC cells. IP with anti-HDAC7 antibody was followed by IB with anti-ubiquitin antibody to detect polyubiquitinated HDAC7. f Representative western blotting of USP10 inhibitor indicating that Spautin-1 reduced the half-life of HDAC7. The NSCLC cells were treated with Spautin-1 at the indicated concentration and time intervals. For graphs in d and f, the relative HDAC7 levels were quantified using Image Lab software. Representative images and statistical analysis of colony formation assay (g), scratch wound assay (h), and transwell assay (i) in A549 cells after treatment with Spautin-1. Migration ability is expressed as the mean scratch area at each time point. The initial scratched area (0 h) was set as 100%. β-actin was used as the internal control. LV, lentivirus. IP, immunoprecipitation. IB, immunoblot