Fig 1: Inhibition of Asm attenuated nicotine-induced podocytes damage. Confocal images represent the expressions of desmin (A) and summarized quantification of desmin (B). Western blot data show the expression of desmin (C) and summarized quantification of desmin (D). N = 15–20 each group for immunofluorescence expression. Ctrl: control, Veh: vehicle, Ami: amitriptyline. Images were quantified using Image J software. * Significant difference from the control; # significant difference from the nicotine-treated group.
Fig 2: Inhibition of Asm attenuated nicotine-induced inflammasomes activation in podocytes. Data are presented as arithmetic means ± SEM (n = 6 per group) for IL-1β production (A) and caspase-1 activity (B) in podocytes exposed to nicotine, with or without amitriptyline, an Asm inhibitor. * Significant difference from the control, # Significant difference from the nicotine-treated group.
Fig 3: Inhibition of Asm protects against nicotine-induced apoptosis in podocytes. Flow cytometry analysis (A) and corresponding quantification (B) were performed to investigate the role of Asm in nicotine-mediated podocyte apoptosis. Data were analyzed using FlowJo v10.10.0 software, with apoptotic cells (%) calculated as the sum of early apoptotic, late apoptotic, and necrotic populations. Results are presented as fold change relative to the control group. * p < 0.05 vs. control group, p < 0.05 vs. nicotine-treated group; # significant difference from the nicotine-treated group. Q1: necrotic cells, Q2: late apoptotic cells, Q3: early apoptotic cells, Q4: live cells, Ctrl: control podocytes, Nico: nicotine (8 µM)-treated podocytes.
Fig 4: Inhibition of Asm attenuated nicotine-induced podocytes damage. Confocal images represent the expressions of Podocin (A) and summarized quantification of Podocin (B). Western Blot data show the expression of Podocin (C) and summarized quantification of Podocin (D). N = 15–20 each group for immunofluorescence expression. Ctrl: control, Veh: vehicle, Ami: amitriptyline. Image analysis was performed using ImageJ software. * Indicates a significant difference from the control group, while # denotes a significant difference from the nicotine-treated group.
Fig 5: Asm inhibition attenuated nicotine-induced ceramide production. Representative immunofluorescence images (A) and quantification analysis (B) depicting ceramide expression in podocytes under different treatment conditions, including nicotine stimulation and/or amitriptyline, an Asm inhibitor. Image quantification was performed using ImageJ software. N = 20. * Indicates a significant difference compared to the control group, while # denotes a significant difference from the nicotine-treated group.
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