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. Author manuscript; available in PMC: 2018 Oct 28.
Published in final edited form as: Nat Med. 2018 Jul 9;24(8):1128–1135. doi: 10.1038/s41591-018-0090-y

Figure 2.

Figure 2

Bleomycin-induced lung fibrosis and protein S-glutathionylation are elevated in Glrx−/− mice, and lowered following transgenic overexpression of Glrx in lung epithelial cells. (a) Collagen in the right superior lobe, 21 days post administration of bleomycin (BLM) WT PBS n = 5, BLM n = 7, Glrx−/− PBS n = 5 & BLM n = 8, pooled from 2 independent experiments. (b) Masson trichrome staining in WT or Glrx−/− mice described in a. Shown are representative images. Scale bar, 50μm. (c) Overall PSSG in lungs from WT or Glrx−/− mice exposed to PBS or bleomycin (n = 5 mice/group). (d) Fas-SSG in lung tissues (WLL) from WT or Glrx−/− mice 21 days post bleomycin. + DTT: negative control, (e) Caspase 3 (Casp3) activity in lungs from WT or Glrx−/− mice 21 days post-bleomycin (n = 5 mice/group) pooled from 2 independent experiments, (f-j) Attenuation of bleomycin- or AdTGFB1-induced fibrosis, PSSG and Casp3 activation in mice overexpressing Glrx in airway epithelia (Epi-Glrx) compared to WT or Ctr groups. Total PSSG (f; n = 5 mice/group), Fas-SSG (g; Top panels), Casp3 activity (h; n = 5/group) or collagen content (I; Sircol, WT: PBS n = 5, BLM n = 8, Ctr PBS n = 4, BLM n = 5; Epi-Glrx n = 5, BLM n = 7 mice/group) in lung tissues. (g) Bottom panels; Assessment of Fas-SSG in Ctr or Epi-Glrx transgenic mice that received AdTGFB1 or AdCtr. Flag-GLRX confirms transgene expression. Representative blots reflect n = 3-4 mice/group. Ctr: CCSP-rtTA single transgenic mice fed dox. * denotes the Flag-Glrx transgene. j: Hydroxyproline in lungs from CCSP-rtTA/TetOP-Glrx bi-transgenic mice following delayed administration of dox. (ctr; n = 5, TGFB1 n = 8 mice/group). Shown are cropped Western blots. *P < 0.05, **P < 0.01, ***P < 0.001, ANOVA, using Tukey’s post-hoc test.