Western blots
Protein expression was detected by Western blots in THP-1 cells, BMDMs, colonic macrophages, colonic epithelial cells, or colon tissues. Briefly, cells or tissues were lysed using RIPA lysis buffer (#P0013B, Beyotime Biotechnology). The cell lysates were centrifuged at 12000 rpm/min for 15 min, and the supernatant was mixed with 5xSDS sample buffer (#P0015L, Beyotime Biotechnology). After boiling, each group of samples was separated by electrophoresis and transferred to PVDF membrane (#FFP39, Beyotime Biotechnology). The membranes were probed with the appropriate antibodies and then detected using Western Blotting Substrate (#180-501, Tanon). The antibodies used are as follows: anti-actin-β (#AF7018, Affinity), anti-GAPDH (#AF7021, Affinity), anti-AKT (#AF6216, Affinity), anti–p-AKT (#AF0016, Affinity), anti-STAT1 (#AF6300, Affinity), anti-p-STAT1 (#AF3300, Affinity), anti-PRDX1 (#DF6652, Affinity), anti-CD1d (#ab215445, Abcam), anti-IL-18 (#ab71495, Abcam), anti-IL-1β (#ab234437, Abcam), anti-ZO1 (#ab216880, Abcam), anti-Claudin1 (#ab180158, Abcam), anti-Occludin (#ab222691, Abcam),anti-PCNA (#13100, CST), anti-p-IκBα (#2859, CST), anti-IκBα (#4812, CST), anti-p-p65 (#3033, CST), anti-p65 (#WL01273b, Wanleibio), anti-ASC (#WL02462, Wanleibio),anti-IL-22 (#WL04441, Wanleibio), anti-IL-10(#sc-365858, Santa Cruz Biotechnology), anti-Caspase-1 (#AG-20B-0042, AdipoGen), and anti-NLRP3 (#AG-20B-0014-C100, AdipoGen). The primary antibody dilution was 1:1000 ~ 1:2000.