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. 2012 Oct 15;186(8):740–751. doi: 10.1164/rccm.201201-0084OC

Figure 3.

Figure 3.

Lactic acid induces myofibroblast differentiation. Primary human lung fibroblasts were cultured with 1, 10, and 20 mM lactic acid for 72 hours. Western blot analysis of protein lysates were performed for markers of myofibroblast differentiation, (A) αSMA and (B) calponin, and compared with cells treated with 5 ng/mL of transforming growth factor (TGF)-β (*analysis of variance [ANOVA], P < 0.05 compared with untreated; n = 3 each). (C) Immunofluorescence for αSMA was performed on cell cultures treated with 10 mM and 20 mM lactic acid and was compared with cells treated with TGF-β (scale bars represent 50 μM). (D) Col1A1 and (E) Col3A1 mRNA induction was analyzed on cells 24 hours after treatment with either 1, 10, or 20 mM lactic acid. Results were compared with untreated cells and cells treated with 5 ng/mL TGF-β (*ANOVA, P < 0.05, n = 3 each). (F, G) Fibroblasts were cultured in the presence of both 20 mM lactic acid and low-dose (1 ng/mL) TGF-β for 72 hours. Protein lysates were analyzed for αSMA expression via Western blot. *ANOVA, P < 0.05; n = 3 each. MEM = Eagle’s minimum essential medium; ns = not significant; Unrx = untreated.