GSDMD N-Terminal Antibody(Mouse specific) - #DF13758

FIGURE 6 Impacts of circPRKAR1B and m6A modification on NLRP3 inflammasome‐mediated pyroptosis. The assessment of the impacts of si‐circPRKAR1B and si‐METTL3 treatment on NLRP3 inflammasome‐mediated pyroptosis using western blotting (A), grayscale analysis of blots (B), immunofluorescence (C; scale bar: 20 μm, white arrows: NLRP3 inflammasome specks and D; scale bar: 25 μm), TUNEL staining (E; scale bar: 25 μm), pyroptosis analysis via SEM (F; scale bar: 100 and 20 μm) and proinflammatory cytokine (IL‐1β/18) level analyses via ELISA (G). METTL3, methyltransferase‐like 3; m6A, N6‐methyladenosine; SEM, scanning electron microscopy; NLRP3, nucleotide‐binding oligomerization domain, leucine‐rich repeat and pyrin domain‐containing 3; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; ELISA, enzyme‐linked immunosorbent assay; IL, interleukin. The data are presented as mean ± SD. ***p < .001 by one‐way ANOVA.

Fig. 1 Irisin inhibited cellular pyroptosis in Min6 cells treated with high glucose. Min6 cells (mouse pancreatic islet β-cells) were incubated with 25 mmol/L glucose for 24 h to construct a T2DM cell model. (A) ELISA was performed to determine the insulin level to evaluate whether the modeling was successful. (B) GSDMD-N immunofluorescence staining was performed to detect cellular pyroptosis after adding 100 ng/mL irisin, followed by incubation for 24 h. (C) Western blotting assays were performed to detect the levels of C-caspase-1 and GSDMD-N. (D) Semi-quantitative analysis of proteins was performed. (E) Caspase-1 activity was detected. (F) ELISA was performed for detecting the level of secretion of IL-1β and IL-18 proteins; ***p 

Fig. 1 Irisin inhibited cellular pyroptosis in Min6 cells treated with high glucose. Min6 cells (mouse pancreatic islet β-cells) were incubated with 25 mmol/L glucose for 24 h to construct a T2DM cell model. (A) ELISA was performed to determine the insulin level to evaluate whether the modeling was successful. (B) GSDMD-N immunofluorescence staining was performed to detect cellular pyroptosis after adding 100 ng/mL irisin, followed by incubation for 24 h. (C) Western blotting assays were performed to detect the levels of C-caspase-1 and GSDMD-N. (D) Semi-quantitative analysis of proteins was performed. (E) Caspase-1 activity was detected. (F) ELISA was performed for detecting the level of secretion of IL-1β and IL-18 proteins; ***p 

Figure 2. Upregulation of miR-140-5p alleviated chondrocyte pyroptosis in OA mice. the OA mice were injected with agomiR-140-5p, with agomiR-NC as the control. A: cleaved caspase-1 positive expression was detected using immunohistochemical staining; the arrow indicates positive staining. B: GSDMD-N protein level was detected using western blot. C: IL-1β and IL-18 levels were detected using ELISA kits. N = 6. the measurement data are expressed as mean ± standard deviation and analyzed using one-way ANOVA, followed by Tukey’s multiple comparisons test, ***p < 0.001