Figure 7
From: Inhibition of apoptotic Bax translocation to the mitochondria is a central function of parkin
K21 and K64 are necessary and sufficient for parkin-regulated Bax translocation to the mitochondria. (a) CHO cells without or with stable expression of parkin (CHO-Parkin) were transiently transfected with untagged wild-type (WT) Bax, Bax with no lysines (ØK Bax) and K21R/K64R Bax. Representative immunocytochemical images of cells showing Bax translocation into the mitochondria (human Bax-specific 2D2 antibody) and after DAPI treatment with 1 μM staurosporine (Sts) for 5 h (b) Quantification of a. Cells were scored visually for Bax translocation into the mitochondria as completely cytoplasmic (open), predominantly cytoplasmic (open dotted), predominantly mitochondrial (gray) or completely mitochondrial (gray dotted) by a blinded observer. Sixty-six to 102 cells were counted per condition. Data plotted from 3 independent experiments. (c) Percentage of pooled mitochondrial fractions (predominantly mitochondrial+completely mitochondrial) plotted from b. All bars represent mean±S.D., n=3. Asterisks indicate significant differences when compared with WT, n.s. indicates not significant (P<0.05)
