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. 2020 Mar 5;21(5):e49248. doi: 10.15252/embr.201949248

Figure 4. Small‐molecule agonist of MCAK activity rescues age‐associated CIN and delays senescence.

Figure 4

  • A
    Mitotic duration of neonatal (N) and elderly (87 years) human dermal fibroblasts (HDF) treated for 24 h with different concentrations of UMK57 (MCAK agonist). n ≥ 58 cells were analyzed per condition. For all subsequent experiments, UMK57 was used at 1 μM for 24 h.
  • B, C
    Representative images (B) and quantification (C) of calcium‐stable k‐fiber intensity levels in metaphase, scored by immunofluorescence analysis of n ≥ 34 tubulin‐stained mitotic cells of neonatal and elderly samples treated with DMSO (−) and UMK57 (+). Levels were normalized to neonatal DMSO‐treated condition. Scale bar, 5 μm.
  • D
    Aneusomy index of chromosomes 7, 12, and 18 measured by interphase FISH analysis of n = cells.
  • E
    Percentage of cytochalasin D‐induced binucleated (BN) n = cells with chromosomes 7, 12, and 18 mis‐segregation (MS).
  • F
    Percentage of micronuclei in n = cells scored when treated with DMSO or UMK57.
  • G
    Percentage of n = cells staining positive for double immunostaining of Cdkn1a/p21 (cell cycle inhibitor) and 53BP1 (≥ 1 foci; DNA damage) senescence biomarkers.
  • H
    Percentage of n = cells staining positive for SA‐β‐galactosidase (SA‐β‐gal) activity.
Data information: All values are mean ± SD of at least two independent experiments. ns P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by two‐tailed (A,C) Mann–Whitney and (D–H) chi‐square tests.