Fig. 1

The Malva parviflora hydroalcoholic extract inhibits LPS-induced NF-κB activity in mouse RAW-Blue macrophages. a RAW-Blue macrophages were untreated or treated with LPS (100 ng/mL) in the presence of the indicated concentrations of M. parviflora hydroalcoholic extract (HE) and 12 h later embryonic alkaline phosphatase (SEAP) activity (driven by NF-κB/AP-1 activation) was determined in the supernatants as described in the “Methods” section. Values are expressed as fold increase relative to SEAP reporter activity in untreated control cells. b RAW-Blue macrophages were pre-treated with LPS or M. parviflora for 30 min and 12 h later embryonic alkaline phosphatase (SEAP) activity (driven by NF-κB/AP-1 activation) was determined in the supernatants as described in the “Methods” section. Data are shown as mean ± SEM. Statistical analysis was performed by two-way ANOVA with repeated measures followed by post hoc Sidak’s multiple comparisons test. This analysis revealed a significant effect for the MpHE concentration F(3,12) = 28.9, p < 0.001; for the LPS administration F(1,4) = 31.49 p = 0.005 and for the M. parviflora and LPS interaction F(3,12) = 30.88, p < 0.001. (***p < 0.001, *p = 0.03). c Cellular viability of RAW-Blue macrophages was evaluated using trypan blue exclusion after exposure to the MpHE at three different concentrations (0.1, 0.5, and 1.0 mg/mL) for 12 h in presence or absence of LPS (100 ng/mL). d RAW-Blue cells were treated with the indicated amount of the M. parviflora in the absence (Ctrl) or in the presence of LPS (100 ng/mL) for 12 h. Cells were fixed and stained with phalloidin and DAPI for F-actin (green) and nuclei (blue) detection, respectively. Images were captured using the Olympus FluoView 1000 confocal multiphoton microscope (Scale bar, 30 μm)