Fig. 3

LCN2 induces the differentiation and secretion of TWEAK in macrophages. A, B Tnfsf12, Tnfrsf12a, and Il6 were detected by RT-qPCR after treating with 0, 10, 50 ng/mL LCN2. 24p3r, Mc4r, inos, and Il10 mRNA in J774A.1 cell were detected by RT-qPCR after treated with 50 ng/mL LCN2. C The intracellular levels of Fn14, TWEAK, p-NFκB P65, p-TRAF2, and TRAF2 proteins were detected by Western blotting after stimulation of J774A.1 cell with LCN2 at concentrations of 0, 10, 50 ng/mL, respectively. D, E TWEAK and CD163 was detected by immunofluorescence in J774A.1 cell after LCN2 treatment. Bar = 100 μm. F Elisa was used to detect the concentration of TWEAK in cell supernatants 24 h after LCN2 stimulation of J774A.1. Data from immunofluorescence assays and Elisa assays are representative of three or more independent experiments are presented as the means ± SEM of triplicate wells. qPCR data are presented as the means ± SEM for triplicate wells from three or more independent experiments. ANOVA was used for comparison between groups: *P < 0.05, **P < 0.01, and ***P < 0.001. ns not significant