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. 2018 Feb 7;9(2):171. doi: 10.1038/s41419-017-0257-3

Fig. 5. NLRP3 inflammasome is involved in nicotine-induced endothelial cells pyroptosis.

Fig. 5

a The expression levels of NLRP3 and ASC were increased in HAECs treated with nicotine (0.1, 1 μM) for 24 h, as determined by western blot results. b The mRNA levels of NLRP3 and ASC were increased in HAECs treated with nicotine (0.1, 1 μM) for 24 h. c–d NLRP3 siRNA silenced the expression of NLRP3 at both protein and mRNA level. e–f The protein levels of Caspase-1, IL-1β, and IL-18 in nicotine-treated HAECs were inhibited after transfection with siNLRP3. NC indicates negative control (n = 4). g The mRNA levels of caspase-1, IL-1β, and IL-18 were inhibited by siNLRP3 in the presence of nicotine (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001. h TUNEL (red) and caspase-1 (green) double-positive cells were decreased in nicotine-treated endothelial cell that transfected with siNLRP3. The nuclei were stained blue with DAPI. Magnification: ×200. Scale bar denotes 100 μm. i Pyroptotic cell death was determined by LDH release, and the relative LDH activity was suppressed by siNLRP3 in the presence of nicotine (n = 3). *P < 0.05. j The percentage of PI positive cells was declined after transfection with siNLRP3 in nicotine-treated HAECs (left: the representative photographs, right: the quantification of PI positive cells). Magnification: ×200. Scale bar = 500 μm. **P < 0.01. The data are represented as mean ± SEM (n = 4)