Fig. 5

NLRP3 inflammasome regulates CSCs formation in SCCHN cell lines. a Western blot analysis of NLRP3 inflammasome in SCCNH cell line and human immortalized oral epithelial cell lysates for comparison. b Caspase-1(P20) and Pro- IL-1β in CAL27 and SCC25 cells were significantly increased after stimulation with LPS(10μg/ml, 6 h) and ATP(5 nM, 1 h) as shown by Western blot analysis. c Production of IL-1β (mature form) from CAL27 and SCC25 cells stimulated with LPS and ATP and treated with MCC950(1 h) as measured by ELISA. Data are expressed as mean ± SEM of three independent experiments performed in triplicate. (d and e) LPS and ATP significantly induced CAL27 cells sphere and colony formation compared with the control group, and MCC950 significantly reduced spheres and colonies; Scale bar = 100 μm. (Mean ± SEM, ***P < 0.001; *P < 0.05). f The expression of NLRP3 inflammasome, BMI1, ALDH1 and CD44 was analyzed by Western blot after treatment with LPS + ATP(6 h) and MCC950(12 h) in CAL27. g CAL27 cells were treated with LPS + ATP(6 h) and 10, 25, and 50 nM MCC950(12 h). The cells were analyzed by immunofluorescence cytochemistry. ALDH1 (green), BMI1 and CD44 (red), DAPI (blue); Scale bar = 50 μm. Data shown are representative of three experiments