Fig. 2: ClpXP protease regulates the turnover of N-module.
From: A salvage pathway maintains highly functional respiratory complex I
a Time-lapse BN-PAGE followed by western blot analysis of CI profiles in wild type (+/+) and CLPP-deficient (−/−) MEFs upon acute respiratory chain disruption via doxycycline-mediated inhibition of mitochondrial protein synthesis (n = 3). b BN-PAGE followed by western blot analysis of CI in wild type (+/+) and CLPP-deficient (−/−) cells upon CRISPR/Cas9 depletion of NDUFB11 subunit (n = 3). c Import of radiolabeled precursors (pre-) of CI subunits into wild type (+/+) and CLPP-deficient (−/−) mitochondria isolated from MEFs (n = 4). d Import of radiolabelled NDUFV2 and NDUFS3 precursors and subsequent incorporation into Complex I in intact heart mitochondria from wild type (+/+) and CLPP-deficient (−/−) animals. After the indicated incubation times mitochondrial complexes were analyzed by BN-PAGE (n = 4). e Pulse labeling (35S-methionine) of mtDNA-encoded subunits in wild type (+/+) and CLPP-deficient (−/−) MEFs followed by chases for indicated time points. Mitochondrial complexes were analyzed by BN-PAGE (n = 3). Antibodies used were raised against proteins indicated in the Figure, with putative CLPP substrates shown in bold. “n” represents number of biologically independent experiments.
