Fig. 8
Lack of endothelial Snrk enhances AngII-induced inflammation by potentiating the NF-κB pathway. Primary GnECs were isolated from Snrkwt/wt/CreVE-Cadh+/− (WT) and Snrkf/f/CreVE-Cadh+/− mice. a Cell lysates of GnECs treated with AngII (1 µM) or a vehicle control (Veh) for 24 h were subjected to western blot analysis of Mcp-1, Tnf-α, Snrk, and Gapdh levels. b GnECs were pretreated with DMSO or an NF-κB inhibitor for 2 h, followed by incubation with AngII or a vehicle control for 24 h. Levels of indicated proteins were determined using western blot and densitometry analysis. n = 6, *p < 0.05 relative to vehicle control; #p < 0.05 relative to GnECs from WT mice; &p < 0.05 relative to DMSO control/AngII. c WT GnECs were treated with AngII for 24 h. The subcellular distribution of Snrk (red) and phosphorylated p-65 (p-p65; green) was determined by double staining (scale bar = 10 µm). d The interaction between SNRK and p-p65 in GnECs in the absence or presence of AngII was assessed using immunoprecipitation and western blot analysis. Input, total cell lysates; IgG, immunoprecipitated with IgG. e–h The effects of Snrk knockdown (e, g) or overexpression (f, h) on luciferase reporter activity from the Tnf-α (e, f) or the Mcp-1 (g, h) promoter. siScr, knockdown control; siSNRK, Snrk knockdown; myc, Myc overexpression vector control; SNRK, Snrk overexpression; pGL3, empty luciferase reporter vector; pGL3-TNF-α, Tnf luciferase reporter; pGL3-MCP-1, Mcp-1 luciferase reporter. e–h, n = 5. (e, g) *p < 0.05 relative to pGL3; #p < 0.05 relative to siScr. (f, h) *p < 0.05 relative to pGL3; #p < 0.05 relative to myc. Student’s two-tailed t-test was used for the statistical analysis. The corresponding source data are available in the Source Data file
