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. 2019 Jul 15;9:10194. doi: 10.1038/s41598-019-46333-2

Figure 4.

Figure 4

Incubation of hepatocytes with 1,5-AF caused accumulation of intracellular AF-AGEs and cell damage. (a) Slot blot (SB) analysis of intracellular AF-AGEs. Cell extracts were prepared from HepG2 cells treated with 0, 5, or 25 mM 1,5-AF for 72 h. The amount of AF-AGEs was calculated from a standard curve prepared with AF-AGEs-BSA. SB analysis was performed three times independently and data are shown as the mean ± S.D. (N = 3). P values were calculated by Tukey’s test or Student’s t-test. **p < 0.01 vs. 0 mM 1,5-AF by Tukey’s test. ++p < 0.01 vs. 0 mM 1,5-AF by Student’s t-test. ##p < 0.01 vs. 5 mM 1,5-AF by Student’s t-test. (b) Cell viability was assessed by the CellTiter-Glo assay. HepG2 cells were incubated for 72 h with 0, 5, or 25 mM 1,5-AF in triplicate. Three independent experiments were performed and data are shown as the mean ± S.D. (N = 3). P values were calculated by Tukey’s test. **p < 0.01 vs. 0 mM 1,5-AF, ++p < 0.01 vs. 0 mM 1,5-AF, ##p < 0.01 vs. 5 mM 1,5-AF.