Extended Data Fig. 4: Glycolytic control of FAs and cell area is independent of ATP.
From: Fructose-1,6-bisphosphate couples glycolytic activity to cell adhesion
a, Loss of PFK, aldolase or GAPDH results in lower ATP levels. Relative ATP levels measured in U-2 OS cells treated with indicated siRNAs. Data were normalized to siCtrl and represent mean ± SEM; n = 5 independent experiments; two-sided One sample t-test (P value (siPFK) = 0.014; (siALDOA) = 0.0024; (siGAPDH) = 0.0136). b-e, Lower ATP levels due to inhibition of oxidative phosphorylation do not affect FA numbers or cell area. b, Representative confocal images of paxillin-labeled FAs in U-2 OS cells treated with DMSO or 1 µM Antimycin A for 48 h. FA segmentation and cell outlines (red) are shown below. c, d, Quantification of FAs per cell and cell area shown in b. Data represent mean ± SEM; n = 3 independent experiments; two-sided unpaired Student’s t-test (P value (c) = 0.3716; P value (d) = 0.8734). e, Relative ATP levels measured in U-2 OS cells treated with DMSO or 1 µM Antimycin A for 48 h. Data were normalized to DMSO and represent mean ± SEM. n = 3 independent experiments; two-sided One sample t-test (P value = 0.0014). f-i, Lower ATP levels due to inhibition of glycolysis phenocopy loss of PFK. f, Representative confocal images of paxillin-labeled FAs in U-2 OS cells treated with PBS or 25 mM 2-deoxy-D-glucose (2-DG) for 48 h. FA segmentation and cell outlines (red) are shown below. g, h, Quantification of FAs per cell and cell area shown in f. Data represent mean ± SEM; n = 3 independent experiments; two-sided unpaired Student’s t-test (P value (g) = 0.002; P value (h) = 0.0069). i, Relative ATP levels measured in U-2 OS cells treated with PBS or 25 mM 2-DG for 48 h. Data were normalized to PBS and represent mean ± SEM; n = 3 independent experiments; two-sided One sample t-test (P value = 0.0216). Ns, not significant; *p < 0.05, **p < 0.01. Scale bars, 25 µm.
