Fig. 2

TLY012 reverses skin fibrosis in bleomycin (BLM)-induced and tight skin-1 (Tsk-1) transgenic mouse models. a Representative double-immunostaining for α-SMA (red), DR5 (green), and nuclei (DAPI, blue) in normal (6w NaCl) and BLM-induced skin fibrosis (3w BLM + 3w NaCl; scale bars, 30 µm). b Relative mRNA DR5 levels in the skin of normal (n = 9) and BLM-induced mice (n = 10). c Representative images of Trichrome-stained sections of control and BLM-induced mice (100 µm). d Analysis of dermal thickness (n = 7 /group, n = 10 6w BLM/PBS), hydroxyproline content (n = 8 /group, n = 9 3w BLM + 3w NaCl/PBS, 6w BLM/PBS), and myofibroblast counts (n = 7 6w NaCl/PBS, T5, 3w BLM + 3w NaCl/T1, n = 9 3w BLM + 3w NaCl/T5, n = 10 3w BLM + 3w NaCl/PBS, 6w BLM/PBS). (e) Measurement of caspase-8 and caspase-3/7 activities (n = 8 6w NaCl/PBS, 3w BLM + 3w NaCl/PBS, n = 10 6w NaCl/T5, n = 7 3w BLM + 3w NaCl/T1, T5). f Representative double-immunostaining for cleaved (Cl.) caspase-3 (red) and α-SMA (green) in BLM-induced mice treated with TLY012 (scale bars, 50 µm). g Relative mRNA in the skin (n = 10 6w NaCl/PBS, T5, n = 8 3w BLM + 3w NaCl/PBS, n = 7 3w BLM + 3w NaCl/T1, T5, n = 10 6w BLM/PBS). h Relative mRNA DR5 levels in the skin of Tsk-1 mice (n = 8 WT, n = 9 Tsk-1). i Representative images of Trichrome-stained sections in Tsk-1 mice (scale bars, 100 µm). j Analysis of hypodermal thickness (n = 8), hydroxyproline content (n = 8 WT/PBS, n = 10 Tsk-1/PBS, TLY012), and myofibroblast counts (n = 8 WT/PBS, Tsk-1/TLY012, n = 10 Tsk-1/PBS). n biologically independent animals. Data are shown as median ± interquartile range. The Mann–Whitney test was used. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs 6w NaCl + PBS or WT + PBS; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001, ^####P < 0.0001 vs 3w BLM + 3w NaCl + PBS or Tsk-1 + PBS