Fig. 2

Wnt10a is upregulated during LR-MSC-to-myofibroblast transition. a, b Mice (n = 6 per group) received either saline or bleomycin (BLM) (5 mg/kg body weight) intratracheally and were killed on day 14. a Myofibroblastic differentiation of lung-resident mesenchymal stem cells (LR-MSCs) in mouse lung tissues was analyzed using a dual immunofluorescence assay to detect the expression of stem cell antigen-1 (Sca-1) and α-smooth muscle actin (α-SMA). b Colocalization of Wnt10a and Sca-1 or α-SMA in mouse lung tissues was determined by immunofluorescence. c–g Mouse LR-MSCs were isolated from the lungs of naive mice using magnetic-activated cell sorting (MACS) and cultured for 7 days. c Cell morphology was revealed by conventional light microscopy. d Cell surface markers were analyzed by flow cytometric analysis. Red line, isotype control; Blue line, respective surface marker staining. e Mouse LR-MSCs were seeded in the appropriate differentiation induction medium. After a 21-day osteogenic or adipogenic differentiation period, cells were stained with alizarin red to assess the accumulation of matrix mineralization or with oil red O to detect the formation of lipid droplets. Mouse LR-MSCs were treated with 10 ng/ml transforming growth factor-β1 (TGF-β1) for 48 h followed by the measurement of the expression of α-SMA and Wnt10a using immunofluorescence (f) and the measurement of the mRNA levels of Wnt10a using qRT-PCR (g). The results are shown as the means ± SD (*P < 0.05 versus control)