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. 2020 Aug 13;11:1881. doi: 10.3389/fmicb.2020.01881

Figure 5.

Figure 5

NLRP3 was necessary for P. gingivalis-, but not S. mitis-induced IL-1β production in THP-1 cells. (A–D) PMA-primed THP-1 cells were transfected with NLRP3 small interfering RNA (siRNA) for 24 (gene detection) or 48 h (protein detection). siRNA-transfected cells were infected with P. gingivalis or S. mitis (MOI = 50) for 2 h. Pro-IL-1β, NLRP3, pro-CASP1, and β-actin in cell lysates were detected by immunoblotting (A). IL-1β mRNA expression was measured by real-time qPCR (B). Cell culture supernatant was collected and assayed for IL-1β (C) and active Caspase-1 (D) secretion by ELISA. (E) PMA-primed THP-1 cells were pre-treated with 10 μm MCC950 and then infected by P. gingivalis of MOI 50 for 2 h. (F) PMA-primed THP-1 cells were pre-treated with 1 and 10 μm MCC950 followed by ATP stimulation for 2 h. (G) PMA-primed THP-1 cells were transfected with NLRP3 siRNA for 48 h and then treated by P. gingivalis or S. mitis (MOI = 50) with or without ATP for 2 h. Both IL-1β (Ga) and active Caspase-1 secretion (Gb) were detected by ELISA. Real-time qPCR data and ELISA represent means ± SD of at least three independent experiments and immunoblot analysis results were representative of at least three experiments. *p < 0.05, **p < 0.01, and ***p < 0.001.