Research Paper Volume 12, Issue 17 pp 17738—17753

Figure 1. Ribes meyeri anthocyanins rescue the senescence phenotype of mouse neural stem cells (mNSCs). (A) Neurospheres clonally derived from 23-month-old (23M-NSC) and 3-week-old (3W-NSC) mouse brains and treated with 100 pg/mL R. meyeri anthocyanins. (B) Differences in neurosphere numbers between 23M-NSC and 3W-NSC. (C) The shape of 3W-NSC neurospheres was more uniform, with an increased number and size (20–50 μm) compared with 23M-NSC neurospheres. (D, E) Treatment with R. meyeri anthocyanins increased the numbers of both 3W-NSC- and 23M-NSC-derived neurospheres compared with controls. (F–H) In 23M-NSCs treated with 100 pg/mL R. meyeri anthocyanins for 48 h, p16^ink4a mRNA expression, cell cycle, and relative telomere lengths were measured. (F) Cell senescence marker p16^ink4a mRNA expression detected by qRT-PCR. (G) Cell cycles were determined using flow cytometry. (H) qRT-PCR detection of relative telomere length. (I) Immunofluorescence staining of TuJ1 and GFAP in 23M-NSCs treated with R. meyeri anthocyanins and control. (J) Quantification of (I). Data are presented as the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.0001 compared with untreated cells.