Fig. 2: Loss of Grx1 in macrophages relieves the inflammation of acute lung injury. | Nature Communications

Fig. 2: Loss of Grx1 in macrophages relieves the inflammation of acute lung injury.

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

Fig. 2: Loss of Grx1 in macrophages relieves the inflammation of acute lung injury.The alternative text for this image may have been generated using AI.

a Flow cytometric analysis of intracellular ROS levels in BMDMs after culture under NOX (n = 3) or HOX (n = 3) for 15 min, P = 0.0004. b GSH/GSSG ratio in BMDMs after culture in NOX (n = 3) or HOX (n = 3) for 2 h, P = 0.0017. c Glrx mRNA expression of AMs from WT mice exposed to NOX (n = 3) or HOX (n = 3) as evaluated by qPCR, P < 0.0001. d Immunofluorescence staining of AMs from WT mice exposed to NOX or HOX (green, Grx1; blue, DAPI; scale bars, 40 μm). e Representative confocal microscopy images of AMs from control or Grx1 KO mice exposed to NOX or HOX stained for S-glutathionylated proteins (green) using Grx1-catalyzed cysteine derivatization (nuclei are stained blue with DAPI; scale bars, 40 μm). f Representative flow cytometry quantification of ROS generation in BMDMs stimulated with LPS (100 ng/mL, 5 min), n = 3 in each group, P < 0.0001. g H&E staining of mouse lung sections 24 h following intratracheal administration of PBS or LPS (5 mg/kg) (scale bars, 100 μm). h Total numbers of leukocytes in BALF from mice 24 h after intratracheal administration of PBS or LPS (5 mg/kg), n = 3, 5, 12, 9, respectively, P(Grx1fl/fl +PBS vs. Grx1fl/fl +LPS) = 0.0008, P(Grx1fl/fl +LPS vs. Grx1fl/fl LysMcre+LPS) = 0.009. i Levels of IL-1β, IL-6, and TNF-α in BALF from mice 24 h after intratracheal administration of PBS or LPS (5 mg/kg) as determined by ELISA, n = 3, 3, 11, 7, respectively, P(Il-1β, Grx1fl/fl +PBS vs. Grx1fl/fl +LPS) = 0.0388, P(Il-1β, Grx1fl/fl +LPS vs. Grx1fl/fl LysMcre+LPS) = 0.018, P(Il-6, Grx1fl/fl +PBS vs. Grx1fl/fl +LPS) = <0.0001, P(Il-6, Grx1fl/fl +LPS vs. Grx1fl/fl LysMcre+LPS) = 0.0002, P(TNFα, Grx1fl/fl +PBS vs. Grx1fl/fl +LPS) <0.0001, P(TNFα, Grx1fl/fl +LPS vs. Grx1fl/fl LysMcre+LPS) =<0.0001. 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, f); one-way ANOVA followed by Tukey’s post-hoc test for (h, i). *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.

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