Journal: The journal of cardiovascular aging

Article Title: GDF11 and aging biology - controversies resolved and pending

doi: 10.20517/jca.2023.23

Figure Lengend Snippet: Summary of the various controversies surrounding GDF11 - status and conclusions

Article Snippet: However, a subsequent paper from David Glass’s lab, then at Novartis, pursuing therapeutics that would antagonize GDF8 and GDF11 to treat age-related muscle dysfunction [ – ] , argued that supplementation with rGDF11 has no effect in aged mice and slows skeletal muscle repair in young mice [ ] .

Techniques: Muscles

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 growth differentiation factor 11 (GDF11) shortens telomere length in Neuro 2a cells. Representative images of telomere quantitative fluorescence in situ hybridization (Telo-FISH, A–D ) in single clone-derived GDF11 KO or wild type (WT) Neuro 2a cells; young (Y, 10 days in culture), old (O, 65 days in culture). Quantification of the average telomere length (E) (two-way ANOVA, interaction: F (1, 217) = 1.513, p = 0.2201; WT vs. GDF11 KO : F (1, 217) = 37.64, p < 0.0001; young vs. old: F (1, 217) = 1.183, p = 0.2779; Sidak’s test, young: WT vs. GDF11 KO , p < 0.0001; old: WT vs. GDF11 KO , p < 0.0018; Y-WT: n = 50, Y-GDF11 KO : n = 68, O-WT: n = 66, O-GDF11 KO : n = 48), and the average relative fluorescence of short telomeres (F) (two-way ANOVA, interaction: F (1, 37) = 6.364, p = 0.0161; WT vs. GDF11 KO : F (1, 37) = 4.643, p = 0.0378; young vs. old: F (1, 37) = 5.642, p = 0.0228; Y-WT: n = 3, Y-GDF11 KO : n = 18, O-WT: n = 8, O-GDF11 KO : n = 12). Data are represented as mean ± SEM, * p < 0.05, ** p < 0.01. Scale bars, 10 μm.

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, Fluorescence, In Situ Hybridization, Derivative Assay

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: Loss of GDF11 regulates telomere maintenance-related genes. Heatmap of upregulated (3, red) and downregulated (4, blue) (A) and bar graphs of 4 downregulated (B) telomere maintenance-related genes caused by deletion of GDF11 in single clone derived Neuro2a cells that were cultured for 65 days (two-tailed unpaired t -test, Tert: t = 7.262, df = 4, p = 0.0019; Rpa1: t = 24.58, df = 4, p < 0.0001; RPA2: t = 15.17, df = 4, p = 0.0001; DCLRE1b: t = 10.63, df = 4, p = 0.0004, n = 3 per group). 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: Derivative Assay, Cell Culture, Two Tailed Test

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

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: Loss of GDF11 enhances SMAD2 binding to the promoters of TERT and TERC. CHIP-qPCR assessment of the enrichment of SMAD2 at the promoters of TERT (A) (two-way ANOVA, interaction: F (1, 12) = 3,595, p < 0.001; WT vs. GDF11 KO : F (1, 12) = 3,176, p < 0.001; IgG vs. SMAD2: F (1, 12) = 4,788, p < 0.001; Sidak’s test, SMAD2: WT vs. GDF11 KO , p < 0.001; four samples per group) and TERC (B) (two-way ANOVA, interaction: F (1, 12) = 101, p < 0.001; WT vs. GDF11 KO : F (1, 12) = 83.6, p < 0.001; IgG vs. SMAD2: F (1, 12) = 157, p < 0.001; Sidak’s test, SMAD2: WT vs. GDF11 KO , p < 0.001; four samples per group) in the single clone-derived GDF11 KO and WT Neuro 2a cells that were cultured for 40 days. Data are represented as mean ± SEM, ** 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: Binding Assay, ChIP-qPCR, Derivative Assay, Cell Culture