Research Paper Volume 14, Issue 19 pp 7662—7691

Figure 5. Lotus germ extract induced DAPK1 expression via the inhibition of age-dependent histone acetylation. (A) Cellular aging suppressed DAPK1 expression and LC3-II expression. NB1RGB cells at several passages were subjected to immunoblot assays using the indicated antibodies. (B) Lotus germ extract induced histone acetylation. Aging NB1RGB cells were treated with the indicated concentration of lotus germ extract for 24 h and subjected to immunoblot assays using the indicated antibodies. (C) Aging NB1RGB cells were treated with the indicated concentration of lotus germ extract for the indicated time and subjected to immunoblot assays using the indicated antibodies. (D and E) Inhibition of histone acetylation increased DAPK1 mRNA expression in aging NB1RGB cells. Cells were treated with DMSO (−) or the indicated concentration of TSA for 24 h and subjected to immunoblotting assay (D) or q-PCR assay (E). (F) The treatment with TSA stimulates autophagosome synthesis. Aging NB1RGB cells were treated with DMSO (−) or 1 μM TSA for 24 h, followed by treatment with or without bafilomycin A1 (1 μg/mL) for 2 h. Cells were subjected to immunoblotting using the indicated antibodies. (G) Lotus germ extract specifically modulated epigenetic control genes in aging cells. Young and aging NB1RGB cells were treated with DMSO (−) or 50 μg/mL lotus germ extract (+) for 24 h and subjected to qPCR. Data are presented as the mean ± SD of three simultaneously performed experiments (E, G). Each P value was calculated using two-way ANOVA; n.s.: not significant, ^*P < 0.05, ^**P < 0.01.