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. 2023 Apr 5;29(1-2):25–34. doi: 10.1177/17534259231166212

Figure 2.

Figure 2

SENP3 facilitates alveolar macrophage M1 polarization and expression of pro-inflammatory genes. Notes: Senp3 fl/fl (n = 6) and Senp3 cKO mice (n = 6) were intratracheally injected with LPS (5 mg/kg) and BALF macrophages were assessed using flow cytometry (A-G). (A) Gate strategy for alveolar macrophages in BALF. (B) CD11b (+) F4/80 (+) cells of the total BALF cells. (C) Frequency of CD11b (+) F4/80 (+) cells. (D) Gated cells were analyzed for CD80 and CD206 expression. Upper left quadrants: CD80 (-) CD206 (+) cells (M2 phenotype); lower right quadrants: CD80(+) CD206 (-) cells (M1 phenotype); upper right quadrants: CD80 (+) CD206 (+) cells (mixed M1/M2 phenotype); lower left quadrants: control. (E) Percentage of CD80 (+) CD206 (-) cells out of all CD11b (+) F4/80 (+) cells. (F) Percentage of CD80 (+) CD206 (+) cells out of all CD11b (+) F4/80 (+) cells. BMDMs were isolated from SENP3 fl/fl (n = 6 per group) and SENP3 cKO mice (n = 6 per group) and stimulated with LPS (100 ng/mL) for 24 h, and the transcription levels of (G) iNOs, (H) IL-6, (I) TNF-α, and (J) IL-10 were measured using qRT-PCR (n = 3 per group). The graphs show the means ± SDs, and the data shown in A–J are representative of three independent experiments. Values of p < 0.05 were considered statistically significant, and data marked with a one (*), two (**) and three (***) asterisks indicate p values of <0.05, < 0.01 and <0.001, respectively. SENP3, SUMO-specific peptidase 3; LPS, lipopolysaccharide; BALF, bronchoalveolar lavage fluid; IL, interleukin; TNFα, tumor necrosis factor α; iNOS, inducible nitric oxide synthase.