Journal: Journal of translational medicine

Article Title: Promoting the transition from pyroptosis to apoptosis in endothelial cells: a novel approach to alleviate methylglyoxal-induced vascular damage.

doi: 10.1186/s12967-025-06195-x

Figure Lengend Snippet: Fig. 4 DT-13 converts MGO-induced pyroptosis to apoptosis by promoting the formation of apoptotic bodies. A. The representative morphology of HUVECs in each group after pretreatment with DT-13 (1 μM) for 6 h and then exposure to MGO (0.8 mM) for 24 h. Scale bar = 50 μm. B. The morphology of dead cells in MGO-induced pyroptosis (top) and in apoptosis following DT-13 pretreatment (bottom). Scale bar = 50 μm. C and D. The number of apop totic bodies in the intercellular space (C) and cell culture medium supernatant (D) was evaluated under MGO induction with and without DT-13. Scale bar = 20 μm. E. Flow cytometry was used to detect the proportion of apoptotic bodies in each group (top), and the proportion of Annexin V and PI staining in apoptotic bodies in each group (bottom). F. Statistical analysis results of apoptotic body content and statistical analysis results of Annexin V-positive/PI- positive in apoptotic bodies (Statistical analysis was performed using one-way ANOVA, followed by Tukey test, n = 3). G. Fluorescence staining of Annexin V (green), PI (red), and DAPI (blue) in pyroptosis cells and apoptotic bodies. The white arrows indicate swollen cell membranes in pyroptotic cells. H. The LDH and IL-18 levels in the supernatants of cell cultures from each group were quantified by LDH Kit and IL-18 ELISA Kit (Statistical analysis was performed using one-way ANOVA, followed by Tukey test, n = 6). Data were presented as mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: DT-13 (1 μM, 130551-41-6, MCE, China) was administered 6 h prior to MGO.

Techniques: Cell Culture, Flow Cytometry, Staining, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Journal of translational medicine

Article Title: Promoting the transition from pyroptosis to apoptosis in endothelial cells: a novel approach to alleviate methylglyoxal-induced vascular damage.

doi: 10.1186/s12967-025-06195-x

Figure Lengend Snippet: Fig. 5 DT-13 promoted cytoskeletal rearrangement and apoptotic body formation via phosphorylation of MLC2. A. Immunofluorescence showed the cytoskeletal morphological changes of HUVECs exposed to different concentrations of MGO (0, 0.2, 0.4, 0.8 and 1.6 mM) for 24 h. Rhodamine-Phalloidin (red) stains the cytoskeleton, DAPI (blue) stains the nucleus. In the negative control group, rhodamine-phalloidin was not used. Scale bar = 20 μm. B. Representative immunofluorescence staining of cytoskeleton in HUVECs after pretreatment with DT-13 (1 µM) for 6 h and exposure to MGO (0.8 mM) for 24 h. Rhodamine-Phalloidin (red) stains the cytoskeleton, DAPI (blue) stains the nucleus (peripheral ring: white arrow). In the negative control group, rhodamine-phalloidin was not used. Scale bar = 20 μm. C. The representative morphological images of HUVECs in different treatment groups (same as B) with or without Ca2+. (Red arrows: pyroptosis cells, yellow arrows: apoptosis cells). Scale bar = 50 μm. D. Effect of the ROCK inhibitor Y27632 (5 µM, 2 h) on the formation of apoptotic bodies induced by DT-13. (Red arrows: pyroptosis cells, yellow arrows: apoptosis cells). Scale bar = 50 μm. E Western blot ting was performed to detect the protein levels of p-MLC2 and MLC2 in different treatment groups, and statistical analysis was performed using Image J (Statistical analysis was performed using one-way ANOVA, followed by Tukey test, n = 3). Data were presented as mean ± SD, *P < 0.05

Article Snippet: DT-13 (1 μM, 130551-41-6, MCE, China) was administered 6 h prior to MGO.

Techniques: Phospho-proteomics, Immunofluorescence, Negative Control, Staining, Western Blot

Journal: Journal of translational medicine

Article Title: Promoting the transition from pyroptosis to apoptosis in endothelial cells: a novel approach to alleviate methylglyoxal-induced vascular damage.

doi: 10.1186/s12967-025-06195-x

Figure Lengend Snippet: Fig. 8 Schematic representation of the mechanism underlying DT-13-mediated inhibition of MGO-induced pyroptosis in HUVECs and promotion of apoptotic body formation. The red line represents the MGO pathway. The black dotted line represents the alternative pathway of MGO upon Caspase3 inhibition. The blue line represents the DT-13 pathway. The Bright blue font represents the action of inhibitors

Article Snippet: DT-13 (1 μM, 130551-41-6, MCE, China) was administered 6 h prior to MGO.

Techniques: Inhibition