Figure 1.

Sfn impedes M1 marker expression during macrophage polarization initiated by LPS. IBMDM macrophages were pretreated with DMSO or the indicated concentrations of Sfn for 30 min before they were stimulated with LPS (25 ng/ml) for 24 h. RNA was extracted, reversely transcribed, and subjected to qPCR analysis for il1β (A), il6 (B), and nos2 (C) mRNA expression. Ppia served as a reference gene; data were referred to as M0 controls. Secretion of TNF-α (D) was assessed using an ELISA, and nitric oxide (E) was measured by monitoring nitrite levels via the Griess assay as described in the Methods section. Levels of intracellular ROS (F) were determined by H₂DCF-DA staining and subsequent flow cytometric analysis. Data depict mean ± SD from at least three independent biological replicates (^* p < 0.05, ^** p < 0.01, ^*** p < 0.001; ANOVA, followed by multiple comparisons test).