Fig. 1: PFOS triggers caspase-1 activation and IL-1β secretion in macrophages.

a, b Bone marrow-derived macrophages (BMDMs) were treated with PFOS in indicated dose points for 6 h, cell lysates were collected to determine mRNA levels of IL-1β, and cell supernatants were collected to measure IL-1β secretion and cell death (a). Immunoblot analysis of of caspase-1 activation and IL-1β maturation in the supernatants, and ASC oligomerization, pro-caspase-1, pro-IL-1β, and GSDMD cleavage in the lysates of PFOS-treated BMDMs (b). c, d PMA-differentiated THP-1 cells (THP-1-derived macrophages) were stimulated with PFOS as indicated for 6 h. Cell lysates were harvested to analyze mRNA levels of IL-1β, and cell supernatants were collected to determine IL-1β and IL-18 production, and cell death (c). Immunoblot analysis of caspase-1 activation and IL-1β maturation in the supernatants, and ASC oligomerization, pro-caspase-1, pro-IL-1β, and GSDMD cleavage in the lysates of PFOS-treated THP-1-derived macrophages (d). e Knockout (KO) efficiency of CASPASE-1 (CAS1) KO and ASC KO THP-1-derived macrophages were evaluated by immunoblot. f, g IL-1β secretion from wild type (WT), two clones of CAS1 KO (KO#1: black plots, KO#2: gray plots) (f) or ASC KO (KO#1: red plots; KO#2: orange plots) (g) THP-1-derived macrophages was determined by ELISA. The cells were treated with PFOS (150 μM, 6 h) or poly (dA:dT) (2 μg/ml, 6 h) or pre-treated with LPS (200 ng/ml, 3 h) followed by ATP (5 mM, 6 h). In a, c, f and g, all error bars, mean values ± SEM, P-values were determined by unpaired two-tailed Student’s t test of n = 3 independent biological experiments. For b, d and e, similar results are obtained for three independent biological experiments. Source data are provided as a Source data file.