Figure 5: Increased glycolytic activation in DJ-1-deficient cells is dependent on AMPK.

(a,b) Immunoblot analysis of phospho-AMPKα(Thr172) protein levels in (a) C2C12 myotubes (n=3 per group) and (b) quadriceps lysates from female mice fed a HFD for 3 months from 2 months of age (n=6 per group). For b, protein bands shown are from non-adjacent lanes on the same gel. Full scan images of immunoblots are shown in Supplementary Fig. 6e,f. (c) Basal ECAR, (d) basal OCR, (e) maximal respiration and (f) coupling efficiency measured using the Seahorse flux analyzer in C2C12 cells co-transfected with Dj1 and Prkaa2 siRNA (n=4 per group). Results are presented as mean±s.e.m. according to the two-tailed unpaired Student’s t-test for (a) and (b), and one-way analysis of variance followed by Tukey’s post-hoc test for c–f. *P<0.05; **P<0.01; ***P<0.001.