Fig. 5
TA-dependent NADPH production supports fatty acid synthesis and ROS scavenging. a, b Relative concentrations of NADPH and fold changes of NADP + /NADPH ratios were detected using a bioluminescent assay (n = 3). c, d Normalized mass spectrometric peak areas of the indicated metabolites. (n = 3) e, f Fraction of M + 2n labeled palmitate, or total IMP, GMP and AMP levels were detected by mass spectrometry-based metabolic profiling (n = 3). g Cellular ROS levels were measured by Cell Rox deep red flow cytometry staining. Mean fluorescence intensity ± SEM was calculated by FlowJo (n = 3). h Percentages of apoptotic cells were detected by Annexin V-FITC and PI staining. Numbers indicate the percentages of total evaluated cells. MDA-MB-361 cells were treated with DMSO or 4 μM lapatinib for 48 h in all assays. Numbers showing percentage of each population ± SEM, P-value shows Q1 and Q2 percentages with student t-test. (n = 3). i, j Relative siTA MDA-MB-361 cell survival with or without 3-day supplementation with NAC (i) or fatty acids (j) combined with DMSO or lapatinib, as indicated (n = 5). *P < 0.01, student t-test, error bars denote the SD
