Figure 4
From: PGC-1β maintains mitochondrial metabolism and restrains inflammatory gene expression
Impaired oxidative metabolism in PGC-1β-deficient DCs is not due to net changes in mitochondrial mass nor is it substrate-specific. DCs were transduced with control shRNA or Ppargc1b shRNA. (a) Mitochondrial mass of DCs stained with MitoSpy Green or MitoTracker Deep Red, with geometric MFIs represented as fold changes relative to the control condition. (b) Protein expression of complexes I-V of the ETC and VDAC of control or PGC-1β-deficient DCs cultured with 0, 1, or 10 mM glucose for 6 h. (c) Gene expression fold changes of Cpt1a (left) and Hadha (right) relative to Hprt. (d) DCs were cultured in nutrient-limiting media for 6 h and OCR was measured immediately following addition of either BSA or BSA-conjugated palmitate. (e) OCR was measured by Seahorse bioanalyzer over time with sequential treatments with oligomycin (oligo), FCCP, and antimycin and rotenone (anti/rot), in assay medium with or without 10 mM glucose or 2 mM glutamine. (f) Basal OCR, coupled respiration, and maximal respiration determined from the experiment in (e). Data from (a,c) are of (a) three and (c) four experiments pooled together, with each individual experiment represented by a circle and conditions from the same experiment joined by a line. Data from (b) is one experiment representative of three experiments. Data from (d–f) are of one experiment representative of two experiments (s.d. of five to six replicates per condition). Statistical significance was determined by (a,c) paired t-test, (d, left) one-way ANOVA, (d, right) unpaired t-test, or (e,g) two-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
