Fig. 6 | Communications Biology

Fig. 6

From: Telomere analysis using 3D fluorescence microscopy suggests mammalian telomere clustering in hTERT-immortalized Hs68 fibroblasts

Fig. 6

Confirmation of hTERT expression and telomere elongation in the generated cell line Hs68 + hTERT. a Immunoprecipitation of Flag-hTERT in Hs68 and Hs68 + hTERT cells followed by western blot for hTERT protein. Low levels of hTERT were detected in input samples of Hs68, but no hTERT was detected after immunoprecipitation with Flag-antibody. In both input and immunoprecipitated samples of Hs68 + hTERT high levels of hTERT were detected, indicating successful infection with hTERT-plasmid. b RNA expression of hTERT in Hs68 and Hs68 + hTERT measured by qPCR. c Telomerase activity of hTERT in Hs68 and Hs68 + hTERT measured by Telomerase Repeated Amplification Protocol (TRAP). d Relative amount of telomeric DNA in Hs68 and Hs68 + hTERT measured by telo-qPCR. Upon hTERT overexpression, there is a 3.5-fold increase in average telomere length in Hs68 + hTERT compared to Hs68 cells. e Absolute telomere length in Hs68 (~5.3 kb) and Hs68 + hTERT (~18.9 kb) measured by Telomere Restriction Fragments (TRF) analysis. This matched the telo-qPCR data and a 3.5-fold increase in telomere length upon hTERT overexpression. Two-tailed unpaired t-tests were performed, n = 3 biologically independent experiments, ****P < 0.0001. Source data used for the graphs in b and d can be found in Supplementary Data 2.

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