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AGEs induce pyroptotic cell death in cardiomyocytes through the RAGE/ROS/NLRP3 pathway. Primary neonatal mouse cardiomyocytes were pretreated with or without FPS‐ZM1 (20 μ M), NAC (10 μ M), MCC950 (10 μ M), or vehicle for 1 h, followed by stimulation with AGEs (200 μ g/mL) for 24 h. (a) Representative fluorescence images of SYTOX green staining. Scale bar, 200 μ m. (b) Quantitative analysis of SYTOX‐positive cells, n = 4. (c) Representative fluorescence images of propidium iodide (PI) uptake. Scale bar, 100 μ m. (d) Quantitative analysis of PI‐positive cells, n = 4. (e) Lactate dehydrogenase (LDH) release in the cell culture supernatant was measured, n = 4.

Journal: Journal of Diabetes Research

Article Title: The AGEs/RAGE Axis Contributes to Myocardial Cell Death in Type 1 Diabetic Cardiomyopathy via the ROS/NF‐ κ B/NLRP3 Inflammasome Pathway

doi: 10.1155/jdr/6438008

Figure Lengend Snippet: AGEs induce pyroptotic cell death in cardiomyocytes through the RAGE/ROS/NLRP3 pathway. Primary neonatal mouse cardiomyocytes were pretreated with or without FPS‐ZM1 (20 μ M), NAC (10 μ M), MCC950 (10 μ M), or vehicle for 1 h, followed by stimulation with AGEs (200 μ g/mL) for 24 h. (a) Representative fluorescence images of SYTOX green staining. Scale bar, 200 μ m. (b) Quantitative analysis of SYTOX‐positive cells, n = 4. (c) Representative fluorescence images of propidium iodide (PI) uptake. Scale bar, 100 μ m. (d) Quantitative analysis of PI‐positive cells, n = 4. (e) Lactate dehydrogenase (LDH) release in the cell culture supernatant was measured, n = 4.

Article Snippet: Trypsin solution (Cat#C0201), CM‐H2DCFDA staining (Cat#S0035S), Cy3‐conjugated secondary antibody (Cat# A0521), FITC‐conjugated secondary antibody (Cat# A0568), 4 ′ ,6‐diamidino‐2‐phenylindole (DAPI) (Cat#C1005), RIPA (Cat#P0013B), protein SDS‐PAGE protein loading buffer (5X) (Cat#P0015), BCA protein concentration measurement kit (Cat#P0011), and lactate dehydrogenase (LDH) cytotoxicity assay kit (Cat#C0016) were purchased from Beyotime (Beijing, China).

Techniques: Fluorescence, Staining, Analysis, Cell Culture