Fig. 3: PPP regulates macrophage phenotype, function and survival.

a, b NADPH/NADP⁺ (a) and GSH/GSSG (b) ratio in untreated, IFN-γ/LPS or IL-4 treated BMDMs were analyzed. c–e G6pdx siRNA transfected BMDMs were stimulated with IFN-γ/LPS for 24 h, NADPH/NADP⁺ (c), GSH/GSSG (d) and ROS (e) were analyzed. MFI, mean fluorescence intensity. f–h Gys1 or Pygl siRNA transfected BMDMs were stimulated with IFN-γ/LPS for 24 h, NADPH/NADP⁺ (f), GSH/GSSG (g) and ROS (h) were analyzed. i Gys1, Pygl or G6pdx siRNA transfected BMDMs were stimulated with IFN-γ/LPS for 24 h, lactic dehydrogenase (LDH) release was analyzed. j BMDMs were pretreated with GPI or 6AN for 30 min prior to stimulation with IFN-γ/LPS ± GSH (0, 1, 5, 10, 20 mM) for 36 h, and LDH release was analyzed. k–m Pygl or G6pdx siRNA transfected BMDMs were stimulated with or without IFN-γ/LPS for 24 or 36 h, Nos2, Tnf, Il6, and Il1b expression was determined by real-time PCR (k), iNOS, TNF and IL-6 expression was determined by western blot (l) and ELISA (m). n Pygl or G6pdx siRNA transfected BMDMs were stimulated with IFN-γ/LPS for 24 h and incubated with E.coli at ratio 1:1 for 3 h, then the E.coli was collected, coated and cultured with ampicillin-resistant agarose solid medium at 37 °C for 36 h, followed by colony-forming units count. Data are presented as mean ± SEM of n = 3 biologically independent experiments (a–d, f, g, j, k, and m) or n = 4 biologically independent experiments (e, h, i and n). P values were calculated using one-way ANOVA.