Fig. 2
Loss of Mst1/2 promotes ageing in macrophages. a RT-qPCR analysis of various Tlrs genes on peritoneal macrophages isolated from Mst1fl/flMst2fl/fl (WT) or Mst1fl/flMst2fl/fl Lyz-Cre (DKO) mice with indicated age. b Proliferation assay of CFSE-labeled CD4+ OT-II T cells co-cultured with WT and DKO BMDMs in presence of OVA protein. c Representative transmission electron micrographs (TEMs) of lipofuscin granules within peritoneal macrophages isolated from 12-month-old WT and DKO mice. ×25 magnification of areas outlined in the main images as indicated. Scale bars, 2 μm. d, e RT-qPCR analysis of Mst1 and Mst2 (d), and immunoblot analysis of Mst1, Mst2 and p-Mob (e) in peritoneal macrophages isolated from WT mice with indicated age. f–h The relative telomere length (T/S ratio) (f), representative fluorescence microscopy images of telomere FISH analysis (red) and nuclei (blue) (g), and relative fluorescence intensity of telomere FISH (h) in peritoneal macrophages isolated from 2-, 8-, or 12-month-old WT and DKO mice. Scale bars, 10 μm. i Relative fluorescence intensities of telomere FISH in peritoneal macrophages isolated from WT and DKO mice with or without NAC supplementation in drinking water for 7 months. ns, not significant (P > 0.05); **P < 0.01, ***P < 0.001, ****P < 0.0001, compared as indicated (Student’s t test). Data are from one experiment representative of three independent experiments with similar results (mean and s.d. of n = 3 (a, d, experimental replicates); n = 5 (f, biological replicates); mean and s.e.m. of n = 50 (h, i, experimental replicates))
