Advanced Glycation End Products Impair Cardiac Atrial Appendage Stem Cells Properties

Figures

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  Figure 1. Rising concentrations of AGEs gradually reduce CASCs proliferation. CASCs were exposed to different AGEs concentrations (50, 100, 200, and 400 µg/mL). Proliferation was measured at different time points: 24 h ((A), n = 18), 48 h ((B), n = 18), and 72 h ((C), n = 21). Data are normalized to BSA and represented as mean ± SEM. * denotes p < 0.05, ** denotes p < 0.01, **** denotes p < 0.0001 vs. BSA.

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  Figure 2. Rising concentrations of AGEs reduced CASCs survival over time. CASCs were exposed to different AGEs concentrations (50, 100, 200, and 400 µg/mL). Survival was measured at different time points: 24 h ((A), n = 28), 48 h ((B), n = 20), and 72 h ((C), n = 25). Data are normalized to BSA and represented as mean ± SEM. * denotes p < 0.05, ** denotes p < 0.01, *** denotes p < 0.001, **** denotes p < 0.0001 vs. BSA.

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  Figure 3. Rising AGEs concentrations increases CASCs apoptosis. CASCs were conditioned with BSA and rising concentrations of AGEs (50, 100, 200, and 400 µg/mL). The percentage of apoptosis was measured over time for 24 ((A), n = 6), 48 ((B), n = 8), and 72 h ((C), n = 7). Data are normalized to the positive control (+, X-VIVO 0% FCS, 2% P/S) and shown as mean ± SEM. * denotes p < 0.05, ** denotes p < 0.01, **** denotes p < 0.0001 vs. BSA.

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  Figure 4. AGEs reduce CASCs migration capacity. Representative examples of CASCs migration when incubated for 72 h with (A) BSA and different AGEs concentrations: (B) 50 µg/mL, (C) 100 µg/mL, (D) 200 µg/mL, and (E) 400 µg/mL. Scale bar = 200 µm. (F) Quantification of CASCs migration after 72 h of incubation with different AGEs concentrations. Fifty µg/mL (n = 6), 100 µg/mL (n = 10), 200 µg/mL (n = 6), and 400 µg/mL AGEs (n = 20). Data are normalized to BSA and shown as mean ± SEM. *** denotes p < 0.001 vs. BSA.

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  Figure 5. CASCs proliferation is improved when blocking RAGE. CASCs were exposed to 400 µg/mL AGEs. In addition, CASCs were pre-incubated with 10 or 25 µM FPS-ZM1. Proliferation was measured at different time points: 24 ((A), n = 13), 48 ((B), n = 12), and 72 h ((C), n = 15). Data are normalized for BSA and represented as mean ± SEM. * denotes p < 0.05 vs. 400 µg/mL AGEs.

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  Figure 6. RAGE inhibition improved CASCs survival. CASCs were exposed to 400 µg/mL AGEs. In addition, CASCs were pre-incubated with 10 or 25 µM FPS-ZM1. Survival was measured at different time points: 24 ((A), n = 14), 48 ((B), n = 12), and 72 h ((C), n = 15). Data are normalized for BSA and represented as mean ± SEM. * denotes p < 0.05 vs. 400 µg/mL AGEs.

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  Figure 7. RAGE inhibition improves CASCs migration. (A) Representative examples of CASCs migration when incubated for 72 h with 400 µg/mL AGEs and pre-incubated with 10 or 25 µM FPS-ZM1. Scale bar = 200 µm. (B) Quantification of CASCs migration (n = 11) when incubated with 400 µg/mL AGEs and pre-incubation with 10 or 25 µM FPS-ZM1 for 72 h. Data are normalized for BSA and represented as mean ± SEM. * denotes p < 0.05 vs. 400 µg/mL AGEs.