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Fig. 1 | Respiratory Research

Fig. 1

From: Cigarette smoke destabilizes NLRP3 protein by promoting its ubiquitination

Fig. 1

Cigarette Smoke Extract (CSE) decreases NLRP3 abundance. a THP1 cells (total 3 × 106 cells) were treated with the indicated concentrations of CSE for 16 h. Cell lysates (Lys) and culture medium (supernatants, Sup) were collected to measure protein levels of NLRP3, NLRP6 (negative control), ASC, Caspase-1, IL-1β, IL-18 and β-actin/GAPDH (loading control) by immunoblotting. Membranes were stripped to detect multiple different proteins. Immunoblot shown is representative of four independent experiments. The lysate blots are from two blots that are loaded at the same time from identical cell lysates. NLRP3, NLRP6 and β-actin are obtained from one, and p45, ASC, and GAPDH are from another blot. For supernatant blots, NLRP3, NLRP6, and p20 are from the same blot, and IL-1β and IL-18 are from a different blot. b THP1 cells were treated with the indicated concentrations of CSE for 16 h before RNA isolation. Shown is the NLRP3 mRNA expression fold changes determined by qRT-PCR in a box plot. Data are representative of four independent experiments. P value was determined by a Kruskal-Wallis test. c Whole lung lysates from non-smoked or smoked C57BL/6 mice were immunoblotted for NLRP3, NLRP6, and β-actin protein. The relative densitometries of NLRP3 protein for the immunoblots are shown in the right panel. P value was determined by a Mann-Whitney test

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