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. 2024 Feb 3;33(9):818–834. doi: 10.1093/hmg/ddae015

Figure 2.

Figure 2

Reactive oxygen species (ROS) levels in TERT/TERC mutants. ROS were detected with the DHE probe in VA-13 cells transfected with pBABE empty vector (transfection control), pBABE-WT TERT/TERC (WT telomerase) or the different TERT and TERC mutants. (A) Representative wide-field microscopy images of single cells are shown for those mutants that presented significant differences compared to pBABE-WT TERT/TERC. In blue, counterstaining of nuclei with DAPI. In red, the DHE probe for ROS detection. (B, C) Nuclear DHE intensity was quantified for TERT (B) and TERC (C) mutants and normalized to pBABE-WT TERT/TERC. The colors of the bars represent the different gene domains in which the mutations are found: yellow controls: pBABE and pBABE-WT TERT/TERC), light green (TEN: p.Leu55Gln to p.Ala202Thr), light blue (RT: pVal694Met to p.Arg865Cys), orange (CTE: p.Val1090Met and p.Thr1101Met) for TERT and pink (P1a stem: n.23G>C), purple (t/PK: n.96_97delCT and n.98G>A), dark blue (CR4/5: n.269G>A and n.325G>T) for TERC. Graph bars represent mean values ± SEM of three independent experiments. The value of each experiment is the mean nuclear DHE intensity of five different microscopy fields (average of 200 cells/experiment). Statistical significance between pBABE-WT TERT/TERC and each TERT or TERC mutant was calculated using two tailed unpaired t-test (*P < 0.05; **P < 0.01; ***P < 0.001).