Journal: Frontiers in Physiology
Article Title: Loss of Growth Differentiation Factor 11 Shortens Telomere Length by Downregulating Telomerase Activity
doi: 10.3389/fphys.2021.726345
Figure Lengend Snippet: In vitro loss of GDF11 downregulates TERT, TERC, and telomerase activity. Quantification of the mRNA level of telomerase reverse transcriptase (TERT, A ) (two-way ANOVA, interaction: F (4, 20) = 0.4232, p = 0.7901; WT vs. GDF11 KO : F (1, 20) = 153.1, p < 0.0001; time: F (4, 20) = 3.983, p = 0.0155; Sidak’s test, 1 day: WT vs. GDF11 KO , p = 0.0007; 10 days: WT vs. GDF11 KO , p = 0.0003; 20 days: WT vs. GDF11 KO , p < 0.0001; 40 days: WT vs. GDF11 KO , p < 0.0001; 65 days: WT vs. GDF11 KO , p < 0.0001; n = 3 per group) and telomerase RNA component (TERC, B ) (two-way ANOVA, interaction: F (4, 20) = 43.49, p < 0.0001; WT vs. GDF11 KO : F (1, 20) = 185.5, p < 0.0001; Time: F (4, 12) = 39.47, p < 0.0001; Sidak’s test, 40 days: WT vs. GDF11 KO , p < 0.0001; 65 days: WT vs. GDF11 KO , p < 0.0001; WT: 1 vs. 40 days, p < 0.0001; WT: 1 vs. 65 days, p < 0.0001; n = 3 per group) as well as the telomerase activity (C) (two-way ANOVA, interaction: F (2, 12) = 5.662, p = 0.0186; WT vs. GDF11 KO : F (1, 12) = 80.51, p < 0.0001; time: F (2, 12) = 9.197, p = 0.0038; Sidak’s test, 1 day: WT vs. GDF11 KO , p = 0.0017; 40 days: WT vs. GDF11 KO , p = 0.0265; 65 days: WT vs. GDF11 KO , p < 0.0001; WT: 1 vs. 65 days, p = 0.0067; GDF11 KO : 1 vs. 40 days, p = 0.0104; n = 3 per group) in single clone-derived GDF11 KO and WT Neuro 2a cells which were cultured for different periods of time. (D) Effect of rGDF11 on the transcription of TERT in WT or GDF11 KO Neuro 2a cells cultured for 65 days (Brown-Forsythe ANOVA test, Brown-Forsythe ANOVA test: F * (DFn, DFd) = 29.64 (2.000, 4.350), p = 0.0029; Welch’s ANOVA test: W (DFn, DFd) = 23.73 (2.000, 3.723), p = 0.0076; Dunnett’s T3 test, WT + BSA vs. GDF11 KO + BSA, p = 0.0109, GDF11 KO + BSA vs. GDF11 KO + rGDF11, p = 0.0159; n = 3 per group). (E) Effect of rGDF11 on the transcription of telomeric RNA component (TERC) in WT or GDF11 KO Neuro 2a cells cultured for 65 days (Brown-Forsythe ANOVA test, Brown-Forsythe ANOVA test: F * (DFn, DFd) = 29.57 (2.000, 3.365), p = 0.0073; Welch’s ANOVA test: W (DFn, DFd) = 18.08 (2.000, 2.697), p = 0.0274; Dunnett’s T3 test, WT + BSA vs. GDF11 KO + BSA, p = 0.0326; GDF11 KO + BSA vs. GDF11 KO + rGDF11, p = 0.7786; n = 3 per group). (F) Effect of rGDF11 on the telomerase activity of WT or GDF11 KO Neuro 2a cells cultured for 65 days (Brown-Forsythe ANOVA test, Brown-Forsythe ANOVA test: F * (DFn, DFd) = 55.10 (2.000, 3.288), p = 0.0030; Welch’s ANOVA test: W (DFn, DFd) = 34.01 (2.000, 3.400), p = 0.0056; Dunnett’s T3 test, WT + BSA vs. GDF11 KO + BSA, p = 0.0151; GDF11 KO + BSA vs. GDF11 KO + rGDF11, p = 0.1590; n = 3 per group). Effects of overexpression of GDF11 (G) on transcription of TERT (H) and TERC (I) in GDF11 KO Neuro 2a cells cultured for 65 days. For statistical analysis details, please see . Data are represented as mean ± SEM, * p < 0.05, ** p < 0.01.
Article Snippet: Recombinant GDF11 protein (rGDF11, Peprotech, Cat#120-11) of 100 ng/ml was added into the DMEM medium (Gibco, Carlsbad, CA, United States) without fetal bovine serum of WT and GDF11 KO Neuro 2a cells at their appropriate cell confluence (>50%), and the treatment lasted for 24 h. Then, the cells were collected for detecting the transcription of TERT and TERC, and telomerase activity.
Techniques: In Vitro, Activity Assay, Reverse Transcription, Derivative Assay, Cell Culture, Over Expression