Fig 2: G-CSF induces pyroptosis through ELANE in leukemia cells.A Representative images of Co-IP using a G-CSF antibody in KG1a cells followed by detection of ELANE protein level by WB. Rabbit IgG was used as negative control. B The mRNA and protein level of ELANE in KG1a cells treated with ELANE siRNAs. C Representative images of flow cytometric analysis for programed cell death in KG1a cells treated with G-CSF or combination of G-CSF and ELANE siRNA. The bar graph showed the quantification of death cell number in each group. D The protein level of N-GSDMD in KG1a cells treated with G-CSF or combination of G-CSF and ELANE siRNA. E The levels of IL-1ß and IL-18 in the cultured medium of KG1a cells treated with G-CSF or combination of G-CSF and ELANE siRNA detected by ELISA. NC: negative control. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 3: Metformin attenuates PM2.5-induced pulmonary oxidative stress and cell death. After PM2.5 exposure, the levels of 3'-nitrotyrosine (3'-NT) (A) and 4-hydroxynonenal (4-HNE) (B) in lung tissue were measured. Lung sections from the control and PM2.5-exposed mice were stained with DAPI (blue) and TUNEL assay kit dye (red) (scale bar = 20 µm, arrows point to TUNEL-positive cells), and the TUNEL-positive cells were quantified (C, D). Lysates of lung tissue were examined by western blotting to determine the expression levels of total and phosphorylated AMPKa, superoxide dismutase 2 (SOD2), peroxiredoxin 3(PRDX3), PRDX5, thioredoxin reductase 2 (TRXR2) and Bcl-2. ß-Tubulin was used as a loading control (E, F). N = 3–5; data are presented as the mean ± SEM; * indicates p < 0.05, ** indicates p < 0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig 4: Metformin attenuates myocardial oxidative stress in PM2.5-exposed mice. After PM2.5 exposure, heart tissue was collected, and the myocardial 3'-nitrotyrosine (3'-NT) (A) and 4-HNE levels were measured. The heart sections were stained with Mitosox (scale bar = 20 µm), and the relative Mitosox fluorescence intensity was quantified (C, D). Lysates of the heart tissue were examined by western blotting for the expression of p-AMPKa, SOD2, PRDX5, PRDX3, TRXR2 and TRX2. ß-Tubulin was used as a loading control (E, F). N = 4; data are presented as the mean ± SEM; * indicates p < 0.05, ** indicates p < 0.01.