Extended Data Fig. 6: Human Th17 cells express the inflammasome components ASC and NLRP3.
From: Human TH17 cells engage gasdermin E pores to release IL-1α on NLRP3 inflammasome activation
a, Lane views of electropherograms obtained with the Jess Simple Western System (ProteinSimple). Monocytes were MACS-isolated with CD14 beads and stimulated in the presence or absence of LPS (24 h) and nigericin (Nig.) for 30 min before cell lysis. Th17 cells were FACS-sorted ex vivo from the blood of healthy donors, stimulated for 48 h and transfected with RNP containing nontargeted control (NTC) or crASC. Cell lysates were collected after 7 days. The experiment was repeated independently 3 times with similar results. b, Western blot analysis. Cell culture lysates derived from Th17 cells and monocytes. Th17 cells were sorted as in (a) and stimulated with anti-CD3 and anti-CD28 mAbs (48 h plate-bound) for 5 days. Monocytes were isolated and stimulated as in (a). The experiment was repeated independently 2 times with similar results. c, NLRP3-speck formation assessed by ImageStream in Th17 cells stimulated for 5 days with anti-CD3 and anti-CD28 mAbs. Each circle indicates an individual healthy blood donor and experiment. Two-sided paired t-test. Data are presented as mean ± SEM. n = 4 biologically independent samples examined over 2 independent experiments.
