Research Paper Volume 4, Issue 12 pp 952—965

Potential anti-aging agents suppress the level of constitutive mTOR- and DNA damage- signaling


Figure 2. Effect of treatment of TK6 cells with different presumed anti-aging drugs for 4 h on the level of constitutive phosphorylation of ribosomal protein S6 (RP-S6).

Exponentially growing TK6 cells were untreated (Ctrl) or treated with the respective agents at concentrations as shown. Phosphorylation status of ribosomal S6 protein was detected immunocytochemically with the phospho-specific Ab (AlexaFluor647), DNA was stained with DAPI; cellular fluorescence was measured by flow cytometry. Top panels: Based on differences in DNA content cells were gated in the respective phases of the cell cycle, as marked by the dashed vertical lines (Ctrl). The percent decrease in mean fluorescence intensity of the treated cells in particular phases of the cell cycle, with respect to the to the same phases of the untreated cells, is shown above the arrows. The dashed skewed lines show the background level, the mean fluorescence intensity of the cells stained with secondary Ab only. Bottom panels: Single parameter frequency histograms showing expression of phosphorylated ribosomal S6 protein (RB-S6P) in all (G1+S+G2M) cells of the respective cultures.