Fig. 4
PITA regulates PFK1 activity and promotes aerobic glycolysis in vivo. a Schematic representation of the transgenic construct used to generate PITA transgenic (TG) mouse lines. PITA protein levels in WT and PITA TG mice at 8 weeks were assessed with both anti-Myc and anti-GAPDH antibodies. b, c Analysis of PFK1, F2,6BP and PK activity in muscle tissues from WT (n = 8) and PITA TG mice (n = 6) maintained on a normal diet. d NADPH levels in muscle of WT (n = 8) and PITA TG mice (n = 6) maintained on a normal diet. e GSG/GSSH (ratio) levels in plasma from WT (n = 8) and PITA TG mice (n = 6) maintained on a normal diet. f Following an overnight fast, WT (n = 8) and PITA TG mice (n = 6) were subjected to a hypereuglycemic clamp. Basal glucose production, whole-body and muscle glucose uptake rate during hypereuglycemic clamp, muscle (soleus and gastrocnemius) glucose uptake during the hypereuglycemic clamp. g, h Serum lactate levels and plasma glucose in WT (n = 8) and PITA TG mice (n = 8) mice in the fasted overnight states. i Glucose tolerance test on WT (opened circle, n = 7) and PITA TG mice (filled circle, n = 7) mice. j Left: Effect of walnut extract on rolling bar test time of WT (n = 7) and PITA (n = 7) mice. Right: Effect of walnut extract on locomotors time of WT and PITA mice. k Body weight of 3-month-old WT (n = 7) and PITA TG (n = 7) mice fed a normal chow. PITA TG mice have similar food intake (n = 7) in comparison with WT mice (n = 7). l Indirect calorimetry of WT (white triangle) and PITA TG mice (black triangle). Oxygen consumption (VO2), carbon dioxide release (VCO2), respiratory exchange ratio (RER; VCO2/VO2) and energy expenditure per kg of body weight were determined in WT and PITA TG mice in metabolic chambers (n = 8 per genotype). m Analysis of glucose consumption and lactate levels, PFK1 activity, NADPH level in CRISPR-Cas9 PITA MEF p53+/+ and MEF p53−/−cells. Data are means ± SD (n=3). Error bars represent mean ± SD for three independent experiments. (mean and s.e.m., n = 3. *P < 0.05: two-tailed unpaired t-test). Unprocessed original scans of blots are shown in Supplementary information, Figure S9. Data in b–l represent mean ± s.e.m. *P < 0.05: two-tailed unpaired t-test
