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. 2017 Oct 5;6(11):2697–2712. doi: 10.1002/cam4.1218

Figure 3.

Figure 3

Ability of RrA to down‐regulate the expression of hTERT rather than to directly affect the activity of telomerase complex. RrA was added into TRAP assay to the final concentration 0.1 U/mL followed by detection of telomerase activity. (A) Telomerase activity determined by TRAP assay. (B) Results of TRAP quantification by densitometry. In order to investigate time‐dependent activity cells were incubated with 0.1 U/mL of RrA followed by detection of hTERT gene expression and protein quantification. To investigate dose‐dependent activity Jurkat and CD4+ T cells were incubated for 9 h with different concentrations of RrA. (C) Time‐dependent and (D) dose‐dependent expression of hTERT measured real‐time RT‐PCR in Jurkat and CD4+ cells. Levels of hTERT expression were normalized relative to the expression of reference gene 18S. (E) Time‐dependent and (F) dose‐dependent changes of hTERT protein amounts measured by western blotting. (G,H) Results of hTERT quantification relative to GAPDH. Data are presented as mean ± SEM. N = 4. P ≤ 0.05 was considered significant by Student's t‐test.