Fig. 1: Grx1-regulated protein S-glutathionylation alleviates hyperoxia-induced acute lung injury. | Nature Communications

Fig. 1: Grx1-regulated protein S-glutathionylation alleviates hyperoxia-induced acute lung injury.

From: Oxidative stress-induced FABP5 S-glutathionylation protects against acute lung injury by suppressing inflammation in macrophages

Fig. 1

a GSH/GSSG ratio in BALF from mice exposed to NOX (n = 3) or HOX (n = 3), P = 0.0003. b mRNA levels of Gss in lung tissue harvested after exposure to NOX (n = 12) or HOX (n = 12) as determined by qPCR, P < 0.0001. c Glrx mRNA expression in lung tissue from mice exposed to NOX (n = 10) or HOX (n = 12), P = 0.0125. d Grx1 activity (reaction mixtures containing L-CySSG) in lung tissue after exposure to NOX (n = 5) or HOX (n = 5), negative control (Grx1 KO mice, n = 3), P(NOX ctr vs. HOX ctr) <0.0001. e H&E staining of mice lungs exposed to NOX or HOX (scale bars, 100 μm). f Morphometric analyses of lung by mean linear intercept (MLI) in all groups as in (e), n = 5 in each group, *** P < 0.0001. g Total numbers of leukocytes in BALF from mice exposed to NOX or HOX, n = 6, 6, 4, 7, respectively, *** P < 0.0001. h mRNA levels of Il1b and Il6 in lung tissue from mice exposed to NOX or HOX as determined by qPCR, n = 5, 3, 5, 5, respectively, P(Il1b, NOX ctr vs. HOX ctr) < 0.0001, P(Il1b, HOX ctr vs. HOX Grx1 KO) = 0.0004, P(Il6, NOX ctr vs. HOX ctr) = 0.0007, P(Il6, HOX ctr vs. HOX Grx1 KO) = 0.0004. i Protein S-glutathionylation in lung tissue from mice exposed to NOX or HOX via non-reducing Western blot (DTT, negative control; β-actin, loading control). j Relative expression of protein S-glutathionylation in lung tissue as in (i). Optical density is normalized by β-actin, n = 3 in each group, ***P < 0.0001. k Protein S-glutathionylation quantified by DTNB assay in lung tissue from mice exposed to NOX or HOX (data shown in nmol GSH/mg protein), n = 35, 30, 25, 25, respectively, P(NOX ctr vs. HOX ctr) = 0.0065, P(HOX ctr vs. HOX Grx1 KO) = 0.0029. l Lung tissue sections from mice exposed to NOX or HOX stained for S-glutathionylated proteins (green). DAPI (blue), Scale bars, 20 μm. All samples were biologically independent and three or more independent experiments were performed. All quantitative data are shown as mean ± SEM and analyzed with a 95% confidence interval. Two-tailed unpaired Student’s t test for (a–c); one-way ANOVA followed by Tukey’s post-hoc test for (d, f, g, h, j, k). *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.

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