Fig. 5

Hsp60 together with CYP11A1 induce mitochondria crista remodeling. A Blue native gel followed by immunoblot detection of CYP11A1 (abbreviated as SCC) and Hsp60 isolated from mitochondria of stable clones of CYP11A1. SCC and Hsp60 complexes appear to form at around 480 to 720 kDa and ~ 900 kDa, respectively. B Coomassie-stained gel of FLAG and HA immunoprecipitation of stable clone C1. MS analysis indicates that the bottom band is CYP11A1 (SCC, labeled by an asterisk), while the top band is Hsp60 (labeled by a circle). C Western blot showing the proteins eluted from the FLAG beads contain CYP11A1 (SCC) and Hsp60 after the induction of C4 cells with doxycycline (doxy). D SCC is present after the C4 lysate is induced by doxycycline (+ doxy) and immunoprecipitated by Hsp60 antibody. T: total fraction, IP: Hsp60 immunoprecipitate. E Western blots showing the depletion of Hsp60 in C4 cells by Hsp60 siRNA (H60). The amount of CYP11A1 (SCC) is reduced after depletion of Hsp60. Scrambled siRNA (sc) and Tim23 are as negative control and loading control, respectively. F Depletion of Hsp60 by siRNA eliminated the induction of mitochondrial tubulovesicular cristae by doxycycline observed under TEM in stable cell clone C4. Unpaired t-test was used, and data shown are mean with standard deviation