Calvin B. Harley, Weimin Liu, Maria Blasco, Elsa Vera, William H. Andrews, Laura A. Briggs, Joseph M. Raffaele. Rejuvenation Research. -Not available-, ahead of print. doi:10.1089/rej.2010.1085.
FIG. 1. Telomerase activation by TA-65® in neonatal foreskin keratinocytes and fetal lung MRC-5 fibroblasts. (A) Keratinocytes in triplicate wells were exposed for 48–72h in different experiments to the dimethylsulfoxide (DMSO) vehicle control, epidermal growth factor (EGF) (positive control, typically 10ng/mL), or TA-65® at indicated concentrations, and products were analyzed as described in Methods. Results from analysis of telomere repeat amplification protocol (TRAP) ladders resolved by gel electrophoresis and quantified by ImageQuant on a PhosphoImager are shown for a typical experiment. (B) MRC-5 cells were exposed to TA-65® at concentrations shown for 48h. Each replicate represents an independent lysate (a replicate culture dish within one experiment). “Chaps” represents the lysis buffer control (no cell extract). HeLa cells are used as positive control cells as described in Methods. T1 is the first telomerase extension product capable of amplification by PCR. IC is the internal control PCR product. Shown is a representative gel from three independent experiments.
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FIG. 2. Baseline telomere length and immune subset data as a function of subject age segregated by cytomegalovirus (CMV) status. Overall baseline data without segregation by CMV status is provided in Table 2. Correlations with age for lymphocyte and granulocute telomere length (A,B) and immune subsets (C–J) at baseline were determined by linear regression in the CMV+and CMV− subjects. Telomere length and immune parameters were analyzed by flow cytometry as described in Methods.
FIG. 4. Relative changes from baseline as a function of time (months) for immune subsets. The mean of the absolute change from baseline for each parameter across all subjects for which data at that time point was calculated and then expressed as a percent change from the mean value at baseline for those subjects. In A–C, data for both cytomegalovirus-positive (CMV+) and CMV− subjects are combined. (D) Relative mean change is shown for the CMV+ population subpopulation only. Asterisks next to a data point signifies p<0.01 (***), p<0.05 (**), and p<0.1 (*) for a two-tailed paired (within-subject)t-test analysis comparing baseline to time point values.