Figure 6
From: Restriction of protein synthesis abolishes senescence features at cellular and organismal levels
Effect of cycloheximide on replicative senescence in normal human fibroblasts.
(a) TIG-7 cells at 67 PDLs were cultured for 1 month to form colonies in the presence and absence of 0.075 μM cycloheximide. Colonies were stained with the senescence-associated ß-galactosidase and Coomassie Brilliant Blue. (b) TIG-7 cells at various PDLs were cultured by serial passages in the presence and absence of 0.075 μM cycloheximide to determine the maximum lifespan. Arrows indicate the time of addition of cycloheximide (n = 4). (c) TIG-7 cells at fully senescent state were cultured in the presence and absence of 0.075 μM cycloheximide for 1 month and the cells were stained with the senescence-associated ß-galactosidase and Coomassie Brilliant Blue. (d) Fully senescent TIG-7 cells were cultured as in (c) and cell growth was monitored (n = 3). (e) Numbers of TIG-7 cells stained with the senescence-associated ß-galactosidase were calculated (n = 3) as in (c). ***P < 0.001 in comparison with the control cells. (f) Fully senescent TIG-7 cells were cultured for 1 month as in (c) and total protein content per cell was determined (n = 3). (g) Mean cell volume for the cells used for (f) (n = 50) was determined. Bars denote means with a standard error of the mean (SEM). *P < 0.05, ***P < 0.001 in comparison between -CHX and + CHX. Scale bars, 100 μm.
