Fig. 5
Genetic inhibition of pY-STAT3 abolishes LPS-challenged hyperinflammation in macrophages. The pY-STAT3 mutant Raw264.7 macrophage cell line was stimulated by LPS (1 μg/ml) for 0.5 or 6 h. a The proliferation of macrophages was analyzed by CCK-8 assay (n = 6 per group; two-tailed Student’s test). b ELISA for IL-6, TNF-α, and TF in macrophage supernatant. c Macrophages TF (red) and nuclei (blue) staining were imaged by laser confocal microscope (scale bar: 1 μm). d The levels of the indicated mRNAs in macrophages were assayed by RT-PCR (n = 4 per group; results are the mean ± SEM; one-way ANOVA test). e Immunoblotting analysis for JAK2/STAT3, NF-κB and MAPK signaling molecules in macrophages (n = 3 per group). **P < 0.01 and *P < 0.05 (control + LPS versus control); ##P < 0.01 and #P < 0.05 (control + LPS versus pY-STAT3mut); ns indicates not significant