Fig. 5

Pyroptosis of B16 cells induced by NSBSO and the mechanism study. A Representative images of the morphology of B16 cells treated with different formulations (10 μM). B Fluorescence images of B16 cells treated with different formulations (10 μM) for 10 h (SYTOX green stained nuclei of dying cells, black arrows indicate cell swelling with big bubbles). C Bio-TEM images of B16 cells at different stages after 10 μM NSBSO treatment and corresponding higher-magnification images of the boxes. D LDH release of B16 cells treated with different formulations for 9 h (Data are presented as mean ± SD, n = 3; ****P < 0.0001). E, F The expression level (E) and semi-quantification analysis (F) of GSDME-N and Cleaved Caspase 3 in B16 cells treated with different concentrations of NSBSO for 6 h (Data are presented as mean ± SD, n = 3; *P < 0.05 and ***P < 0.001). G Extracellular ratio of ATP of B16 cells treated with different formulations for 9 h (Data are presented as mean ± SD, n = 3; ***P < 0.001 and ****P < 0.0001). H Immunofluorescence images of ecto-CRT in B16 cells treated with different formulations (10 μM) for 10 h