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. 2019 Jun 11;10:1291. doi: 10.3389/fimmu.2019.01291

Figure 4.

Figure 4

Flagellin induces NLRC4+ and NLRP3+ specks formation in MDDC. 2 × 105 MDDC from healthy donors (HD-DC; n = 3) or HIV-infected patients (HIV-DC; n = 3) were cultured in 16-wells chamber slides and stimulated with 5 μg/mL FLG or 1 μg/mL LPS for 24 hours at 37°C 5% CO2 with or without 1 mM ATP for more 15 minutes. Mouse anti-human NLRP3 and rabbit anti-human NLRC4 antibodies and fluorescent secondary antibodies (Alexa 488-conjugated goat-anti-mouse IgG1; Alexa 647-conjugated goat-anti-rabbit IgG1) were used to label NLRP3+ and NLRC4+ specks, respectively. DAPI was used to counterstain nuclei. Images acquisition was performed using a DMi8 confocal laser scanning microscope. A representative experiment (magnification: 63x) was reported for HD-DC (A) and HIV-DC (B). Arrows indicated NLRC4+ or NLRP3+ specks. (C,D) CTCF index for NLRP3+ and NLRC4+ specks in untreated (UN) and FLG-treated (FLG) MDDC. Data are represented as mean ± standard error. Mann-Whitney test was applied to compare UN and FLG conditions in (C,D) (*p < 0.05).