Figure 2.

TGFβ decreases pyruvate kinase activity and increases serine-glycine metabolism. PKM2 activator decreases serine-glycine metabolism in myofibroblasts. (A & H) Hydroxyproline in LX2 (A) and NFL (H) with (filled bars) or without (open bars) TGFβ treatment and treated with DASA-10 (grey bars) after TGFβ treatment was analyzed using hydroxyproline kit. (B, C, I & J) Glycolytic intermediates G6P (B, I) and 2-PG (C, J) in LX2 (B, C) and NFL (I, J) with (black bar) or without (open bar) TGFβ treatment and treated with DASA-10 after TGFβ treatment (grey bar) were analyzed using commercial kits. The G6P is presented as μM (1x10⁶ cells lysate in 100 μL), 2-PG is presented as nmole per μL cell lysate. (D, K) Pyruvate kinase activity of lysate of LX2 (E) and NFL (J) with (black bar) or without (open bar) TGFβ treatment and treated with DASA-10 after TGFβ treatment (grey bar) was analyzed using pyruvate kinase activity kit. Pyruvate kinase activity is presented as mU/mL cell lysate. (E, F, L & M) Levels of serine (E, L) and glycine (F, M) in lysate of LX2 (E, F) and NFL (L, M) with (black bar) or without (open bar) TGFβ treatment and treated with DASA-10 after TGFβ treatment (grey bar) was analyzed using amino acid analysis kits. (G) Hydroxyproline in LX2 with or without TGFβ treatment and with or without DASA-10 treatment after TGFβ treatment (as indicated) was analyzed using hydroxyproline kit. The hydroxyproline in A, G, and H is presented as μg of hydroxyproline in lysate of 1x10⁶ cells. Error bars in all panels represent mean ± S.E.M.