Fig. 3

7E protected macrophages from hyperosmotic stress-induced activation. a–d Murine bone marrow cells were isolated for cell culture and treated with M-CSF (100 ng/ml) for 5 days to differentiate into BMDMs. Cells were further treated with 40 mM or 80 mM NaCl to mimic dry eye conditions. Drugs including Xiidra and 7E were added into the medium for 24 h, PBS and mIgG were used as control groups. Cells were harvested and analyzed for the expression levels of several proinflammatory factors including Nos2, Il1b, Tnfa, and Il6. Gapdh was used as an internal control. One-way ANOVA, **p < 0.01, ***p < 0.001 and ****p < 0.0001. Data are shown as the mean ± SEM. e Murine bone marrow-derived M0-type macrophages were treated LPS + IFN-γ (M1), 80 mM NaCl + PBS, 80 mM NaCl + Xiidra, 80 mM NaCl + mIgG, 80 mM NaCl + 7E for 24 h and analyzed for expression of F4/80, CD86, and CD206 by flow-cytometry. The experiments in a–d were repeated three times independently with similar results, and the data of one representative experiment was shown. M-CSF macrophage colony-stimulating factor, BMDMs bone marrow-derived macrophages, LPS lipopolysaccharide, IFN-γ interferon-γ