Fig. 6

Inhibition of PGK1 mitigates M1 macrophages and reduces pyroptosis A-B: The small interfering RNA (siRNA) was employed to downregulate PGK1 expression in BMDMs. Western blotting analyses the expression of PGK1fig in BMDMs. C-E: To simulate M1 macrophages, BMDMs were exposed to LPS for a duration of 24 h. Prior to the stimulation, siRNA was administered. Co-IP assays were performed to detect the phosphorylation level of NLRP3 in BMDMs(C).CD86 expression on BMDMs was detected by flow cytometry after gating with the macrophage marker F4/80. MFI analysis was conducted to compare the expression of CD86 in BMDMs with the siNC group(D). Immunofluorescence analysis of iNOS(Red) staining(scale bar 50 μm) (E). F-I: The cells underwent pre-stimulation with LPS for a duration of 4 h. Afterward, the cells were activated with ATP to induce the pyroptosis model. Protein levels of GSDMD in BMDMs were determined by Western blotting(F-H). Immunofluorescence staining for the colocalization of NLRP3(Green) and Cleaved-caspase1(Red) (scale bar 100 μm) (I). The data shown are the means ± SEM obtained from three separate biological replicates. * P < 0.05, ** P < 0.01 by unpaired t test