Figure 8.

Mitochondrial recruitment of VCP depends on the UBA and UBX domain of UBXN1. (A) HeLa cells transfected with expression plasmids for mitoYFP-T2A-MYC-PRKN, FLAG-UBXN1 (not shown), FLAG-UBXN1ΔUBX or FLAG-UBXN1ΔUBA and treated with 25 µM CCCP for 90 min were fixed and stained using mouse anti-VCP and rabbit anti-FLAG antibodies. Shown are representative experiments of three independent experiments. Scale bar: 20 µm. (B) Colocalization of mitoYFP and VCP or (C) mitoYFP and FLAG-UBXN1, FLAG-UBXN1ΔUBX or FLAG-UBXN1ΔUBA was quantified by Pearson’s correlation in confocal images of A. The boxplots represent three independent experiments with at least 30 cells per condition and experiment. Statistical significance was assessed by one-way ANOVA followed by Student’s t-test with “fdr” adjustment for multiple testing. n.s. – p > 0.05; *** – p < 0.001.