Supplementary Figure 4: GSC bioenergetics is downregulated in AMPK-silenced GSCs.

a, b, Molecular network of most downregulated glycolysis genes (a) and mitochondrial genes (b) in two AMPKβ1-sienced GSCs. Node colour denotes extent of downregulation (darker = greater downregulation). Ingenuity Pathway was also used to confirm NetWalker analysis. c, Heatmap showing differentially expressed genes in indicated pathways in AMPKβ1 shRNA GSCs relative to NT shRNA expressing GSCs. d. Q-RTPCR analysis of HIF1α, GABPA and their target genes in GSCs using a second AMPKβ1 shRNA (Average of two independent experiments). ⁺p = 0.001; * 0.01; ^$* 0.002; ^# 0.006; ** 0.003; ^$0.02; ⁺* 0.05. e, WB of indicated proteins in GSCs expressing NT or AMPKβ1shRNA. f, Glucose uptake (at 30 min) in GSCs expressing NT or AMPKβ1shRNA. . (n = 3). g, WB of GLUT3 in GSCs expressing NT or AMPKβ1shRNA. h, WB of PGC1α in nontarget and AMPKβ1 shRNA expressing GSCs. i, WB using anti-acetyl antibody following immunoprecipitation with PGC1α antibody in control (NT) and AMPK silenced GSC. j, WB showing overexpression of Myc-tagged PGC1α in GSCs. k, Q-PCR analysis of ND4 and β actin DNA in control and AMPK-silenced GSCs overexpressing PGC1α. (Average of two independent experiments). ns = nonsignificant. l, Oxygen consumption rate (OCR) in control and AMPK-silenced GSCs overexpressing PGC1α. (Average of two independent experiments). m, Q-RT-PCR analysis of ND4 and β actin RNA in control and AMPK-silenced GSCs treated with SIRT1720 for 48h. (Average of two independent experiments). n, OCR in control and AMPK-silenced GSCs treated with SIRT1720. (Average of two independent experiments). Error bars; mean +/- S.D. Statistical significance in above experiments was assessed using two-tailed t-test. n values represent independent experiments. Source data are available in Supplementary Table 4. All western blots represent data from 2 (e, h, I, j) or 3 (e, g) independent repeats. Unprocessed blots are shown in Supplementary Fig. 9.