Fig. 1: GPR50 is transiently recruited to the mitochondrial region upon mitochondrial membrane depolarization.
From: GPR50 regulates neuronal development as a mitophagy receptor
A–D Western blotting analysis of endogenous GPR50, TOMM20, and LC3 II levels in the whole cell lysate and mitochondria fractions of Hela cells treated with CCCP for a distinct time as indicated. B Relative levels of TOMM20 in the whole cell lysate. One-way ANOVA (F3, 16 = 13.769, p = 0.000) followed by Dunnett T3 post hoc tests for CCCP 0 h versus 2 h (p = 0.025), 0 h versus 6 h (p = 0.045), 0 h versus 12 h (p = 0.026). n = 5 independent experiments (B). C, D Relative levels of GPR50 and LC3-II in the whole cell lysate and mitochondria fractions. One-way ANOVA for Mitochondria fractions (F3, 16 = 9.110, p = 0.001) followed by Dunnett T3 post hoc tests for 0 h versus 2 h (p = 0.042), 0 h versus 6 h (p = 0.045). One-way ANOVA for whole cell lysate (F3, 16 = 0.960, p = 0.436) followed by Dunnett T3 post hoc tests for CCCP 0 h versus 2 h (p = 0.998), 0 h versus 6 h (p = 0.581), 0 h versus 12 h (p = 1.000) (C). One-way ANOVA for Mitochondria fractions (F3, 16 = 20.603, p = 0.000) followed by Dunnett T3 post hoc tests for 0 h versus 2 h (p = 0.008), 0 h versus 6 h (p = 0.010). One-way ANOVA for whole cell lysate (F3, 16 = 9.132, p = 0.001) followed by Dunnett T3 post hoc tests for 0 h versus 2 h (p = 0.004) (D). n = 5 independent experiments (C, D). E, F HeLa cells transfected with GPR50-FLAG WT were treated with CCCP for the indicated time and immunostained for FLAG, TOMM20, and DAPI (E). Scale bars: 5 μm. F Colocalization coefficient between GPR50 and TOMM20. One-way ANOVA (F6, 186 = 19.451, p = 0.000) followed by Dunnett T3 post hoc tests for CCCP 2 h versus 6 h (p = 0.019), 6 h versus 24 h (p = 0.000), 6 h versus 36 h (p = 0.000), 6 h versus 48 h (p = 0.000). n = 16–34 cells per group from 3 independent experiments (F). Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001. n.s, non-significance.
