Figure 5.

Mitochondrial respiration recovery, DNA repair, and survival after mitochondrial shutdown are compromised by inhibition of glycolysis

(A and B) Log2-values of changes to measured basal respiration (A) and ATP production (B) induced by combination treatment with 2DG (20 mM) and IR with a single dose of 3 Gy compared with non-irradiated controls 1, 6, and 24 h after treatment.

(C and D) Log2-values of changes to measured basal respiration (C) and ATP production (D) induced by fractionated dose of 3 × 3 Gy applied every 24 h for 3 consecutive days compared with non-irradiated controls 1, 6, and 24 h after the last IR dose. Mean values of n = 8–16 wells per cell line from N = 2 independent experiments are indicated. p values were calculated using two-way ANOVA with Tukey’s multiple comparisons post hoc test. All metabolic parameters were normalized to Hoechst intensity (relative fluorescence units, RFU) in each well. OCR, oxygen consumption rate; FC, fold change.

(E) Representative pictures of nuclear γH2A.X on NCI-H60 and MDA-MB-231 cell lines treated with 2DG (20mM) or Rotenone (0.8 μM) in combination with IR (3 Gy) at the indicated time points. Blue, Hoechst 33342; red, γH2A.X. Scale bar: 2 μm.

(F) Heatmap representing fold change (FC) values of measured γH2A.X foci number per area normalized to 0.5 h time point after treatment with IR (3 Gy) alone or in combination with 2DG (20 mM) or Rotenone (0.8 μM) at 6 and 24 h. For corresponding DNA repair kinetics determined using the γH2A.X assay with statistical test refer to Figure S9D. See also Figures S8 and S9.