Fig. 5

GSK461364 suppresses the proliferation of fibroblasts but not that of lung epithelial cells in vivo. C57BL/6 mice received bleomycin (3 mg/kg, intratracheal instillation) on day 0. BI2536 (20 mg/kg, injected intraperitoneally) or vehicle was administered twice a week. The lung tissue was harvested on day 21. Paraffin-embedded lung sections were stained with an anti-pro-SPC antibody (green) and anti-Ki-67 antibody (red) or with an anti-α-SMA antibody (green) and anti-Ki-67 antibody (red). Representative images of immunofluorescence staining in each group are shown (A, E: control group; B, F: bleomycin-alone group; C, G: bleomycin and BI2536 20 mg/kg group). Yellow arrows indicate cells double-positive for α-SMA and Ki-67 (A-C), for pro-SPC and Ki-67 (E-G). Scale bars, 100 μm. (D) The α-SMA⁺ Ki-67⁺cells were counted in 10 random fields using 3 lung sections. Data are displayed as dot plots and the mean. (H) The pro-SPC⁺Ki-67⁺cells were counted in 10 random fields using 3 lung sections. Data are displayed as dot plots and the mean. C57BL/6 mice received bleomycin (3 mg/kg, intratracheal instillation) on day 0. GSK461364 (5 mg/kg, injected intraperitoneally) or vehicle was administered twice a week. The lung tissue was harvested on day 21. Paraffin-embedded lung sections were stained with an anti-pro-SPC antibody (green) and anti-Ki-67 antibody (red) or with an anti-α-SMA antibody (green) and anti-Ki-67 antibody (red). Representative images of immunofluorescence staining in each group are shown (I, M: control group; J, N: bleomycin-alone group; K, O: bleomycin and GSK461364 5 mg/kg group). Yellow arrows indicate cells double-positive for α-SMA and Ki-67 (I-K), for pro-SPC and Ki-67 (M-O). Scale bars, 100 μm. Data were analyzed using a one-way analysis of variance followed by Dunnett’s multiple comparison test and displayed as dot plots and the mean ± SD. For all graphs: NS = not significant; *** P < 0.0001. The P values of each experiment are shown above each figure