Figure 6.

Pin1 knockdown down-regulated NLRP3 inflammasome in human septic shock. (A) Analysis of inflammatory genes in published clinical datasets GSE26440. (B) PMA-differentiated THP-1 cells were transfected with control siRNA or siPin1 followed by stimulation with 500 ng/ml lipopolysaccharide (LPS) for 4 h. Cell extracts were immunoblotted for NLRP3 inflammasomes. (C) PMA-differentiated THP-1 cells were transfected with control siRNA or siPin1 followed by stimulation with 500 ng/ml LPS for 4 h and treatment with 5 mM ATP or 20 mM nigericin for 30 min. Immunoblot was performed for caspase-1 activation. in cell extracts. (D) PMA-differentiated THP-1 cells were transfected with control siRNA or siPin1 followed by stimulation with 500 ng/ml LPS for 4 h and treatment with 5 mM ATP or 20 mM nigericin for 30 min. Immunoblot was performed for GSDMD cleavage in cell extracts. (E, F) PMA-differentiated THP-1 cells were transfected with control siRNA or siPin1 followed by stimulation with 500 ng/ml LPS for 4 h and treatment with 5 mM ATP or 20 mM nigericin for 30 min. Supernatants were analyzed for LDH, IL-1β, and IL-18. (G) Immunoblot analysis for activation of p38 MAPK in PMA-differentiated THP-1 cells transfected with control siRNA or siPin1 and stimulated with LPS (500 ng/ml) for 0, 10, 20, and 30 min. The data represent the mean ± SD of one among three biological replicates, with three technical replicates each. *P < 0.05; **P < 0.01; ns, no significance; two-way ANOVA (E, F).