Induction of NLRP3 Inflammasome Activation by Heme in Human Endothelial Cells

Figure 3

Heme induces NLRP3 expression and activation of caspase-1. (a) LPS-primed (10 μg/mL, 24 h) or nonprimed HUVECs were exposed to heme (10, 25, and 50 μmol/L in 1% FBS, 4 h). NLRP3 mRNA levels were determined by qRT-PCR. (b) LPS-primed (10 μg/mL, 24 h) or nonprimed HUVECs were exposed to heme (25 μmol/L in 1% FBS, 6 h). NLRP3 were analyzed by Western blot from whole cell lysate. Membrane was reprobed for GAPDH. Representative blots of 2 independent experiments are shown. (c) Densitometric analysis of Western blots. (d) C57BL/6 and Nlrp3^−/− mice were injected with heme (300 nmol/peritoneal cavity) or vehicle (Ctrl). Protein expressions of activated caspase-1 and processed IL-1β were analyzed by Western blot from liver samples (16 h). Membrane was reprobed for GAPDH. Representative blots of 3 independent experiments are shown. (e and f) Densitometric analysis of Western blots. Results are shown as mean ± SD of 3 independent experiments. , , and .

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