Fig. 4

Effects of the S. horneri 70% EtOH extract and its CH₂Cl₂- and water-soluble subfractions on (a) the activated NF-κB-p65 expressions, (b) NF-κB DNA-binding activity in LPS-stimulated RAW264.7 cells. Immunofluorescence assay for the nuclear translocation of NF-κB p65. The DAPI-stained or non-stimulated RAW264.7 group displayed blue color and no intense green fluorescence in the nucleus. In lipopolysaccharide (LPS) stimulated cells with p65 staining displayed intense green fluorescence in the nucleous. The S. horneri (c) 70% EtOH extract, (d) CH₂Cl₂-soluble, and (e) water-soluble fractions decreased nuclear localization of NF-κB p65 and reduced green fluorescence intensity within the nucleus in LPS-treated RAW264.7 cells; quantification of NF-κB p65 nuclear localization is displayed for (f) 70% EtOH extract, (g) CH₂Cl₂-soluble, and (h) water-soluble fractions. The white arrows indicate the expression of p65 with intense green fluorescence. Immunoblots of the detected NF-κB-p65 expressions was quantified using ImageJ software, and specific band intensity was normalized to DAPI. Data are expressed as means ± SD (n = 4). *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the LPS-stimulated groups.