Fig. 6: miR-147b reduces CIV activity but not membrane potential and ROS production.

a CIV activity of permeabilized A549 cells transfected with miR-control, miR-147b mimic, or siNDUFA4 as measured by Seahorse. Data were presented as mean ± SEM; P = one-way ANOVA; n = 8 technical replicates per condition. b Seahorse uncoupled electron flow analysis of CIV activity in permeabilized A549 cells transfected with miR-control, miR-147b mimic, or siNDUFA4, as shown in (a). Oxygen consumption rate (OCR) reads were normalized with protein levels quantified with BCA. Dotted line shows the compounds injected at each time point. CIV activity was measured as activity between the maximal activity after TMPD addition and the minimal activity after azide injection. AA antimycin A, TMPD N,N,N′,N′-Tetramethyl-p-phenylenediamine dihydrochloride, Oligo oligomycin. Data were presented as mean and SEM of eight biological replicates. c CIV activity of permeabilized WT or ATGmut-mRNA overexpressing- HAECs and A549 cells, as measured by Seahorse. Data were presented as mean ± SEM; P = two-tailed unpaired Student’s t test; n = 8 technical replicates per condition. d Membrane potential of HAECs and A549 cells with indicated transgenes as measured by flow cytometry of TMRE dye incorporation. Data were presented as mean ±  SEM; P = one-way ANOVA; n = 4 biological replicates per condition from two repeats. e Total cellular ROS levels of A549 cells with indicated transgenes as measured by Amplex Red assay. Data were presented as mean ± SEM; P = one-way ANOVA; n = 4 biological replicates per condition. Data for control and MOCCI were shown in Fig. 5g. f Total cellular ROS levels of HAECs and A549 cells with indicated transgene as measured by flow cytometry of DCF dye reaction. Data were presented as mean ± SEM; P = one-way ANOVA; n = 3 biological replicates per condition. g Effects of MOCCI, miR-147b mimic, siNDUFA4, and ATGmut-mRNA on CIV activity, membrane potential (MP), and ROS production.