Journal: Science Advances

Article Title: circHERC1—A telomerase activator

doi: 10.1126/sciadv.adz3680

Figure Lengend Snippet: ( A ) TRAP analysis of P3 and P12 HUVECs for telomerase activity (32 PCR circles). ( B ) Telomere length analysis of HUVECs at P3 to P12. ( C ) Telomere length–circHERC1 correlation in PBMCs of healthy donors. ( D ) Correlation of circHERC1 and TERT expression in PBMCs (healthy donors; CB1 site mutant). ( E ) Telomere length–circHERC1 correlation in PBMCs (healthy donors; CB1 site mutant). ( F ) TRAP analysis of HUVECs after circHERC1 overexpression and knockdown (32 PCR circles). ( G ) Telomere length in HUVECs after circHERC1 overexpression and knockdown. ( H ) Western blot of TERT in Dox-treated circHERC1-modified HeLa cells. ( I ) TRAP analysis of HeLa cells treated with Dox or not (28 PCR circles). ( J ) TRAP analysis of HeLa cells treated with H 2 O 2 or not (28 PCR circles). ( K )Telomere length of HeLa cells following circHERC1 overexpression and knockdown under conditions of DNA damage or oxidative stress. ( L ) TRAP analysis of 293FT cells transfected with circHERC1 and TB mutant (27 PCR circles). ( M ) TRAP analysis of 293FT cells transfected with circHERC1 and PB mutant (29 PCR circles). ( N ) Telomere length of 293FT cells transfected with circHERC1 and TB mutant. ( O ) Telomere length of 293FT cells transfected with circHERC1 and PB mutant. ( P ) Replicative life span of HUVECs with or without circHERC1 transfection. ( Q ) SA-β-gal staining of HUVECs after circHERC1 overexpression and knockdown. ( R ) Western blot of p21 and p53 in HUVECs after circHERC1 overexpression and knockdown. ( S ) Western blot of phospho-H2A.X in HUVECs after circHERC1 overexpression and knockdown. ( T ) SA-β-gal staining of HeLa cells transfected with circHERC1 and its mutants (Dox-treated). ( U ) Western blot of p21 and p53 in HeLa cells transfected with circHERC1 and circHERC1 PB1 and PB2 mutant. ( V ) Western blot of p21 and p53 in HeLa cells transfected with circHERC1 and circHERC1 TB1, TB3, and TB7 mutant. N values are shown within the panels. All values are means ± SEM.* P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: The TRAP assay was conducted utilizing the TRAPeze® Telomerase Detection Kit (Merck, USA) following the manufacturer’s protocol ( ).

Techniques: Activity Assay, Expressing, Mutagenesis, Over Expression, Knockdown, Western Blot, Modification, Transfection, Staining

Journal: Frontiers in Aging

Article Title: The truncated isoform of the receptor for hyaluronan-mediated motility (RHAMM Δ163 ) modulates shelterin and telomerase reverse transcriptase transcription affecting telomerase activity

doi: 10.3389/fragi.2025.1604051

Figure Lengend Snippet: RHAMM Δ163 regulates telomerase-mediated telomere elongation. RHAMM is present in multiple isoforms, including RHAMM FL and RHAMM Δ163 in mice (A) . HMMR was eliminated using CRISPR-Cas9 in MDA-MB-231 cells. RNA was isolated, and qRT-PCR was used to measure the mRNA expression of TERT, TPP1 , and POT1 , normalized to Gapdh . Data are means ± SD; n = 3 experiments; *p < 0.05, **p < 0.01, ***p < 0.001 by Student’s t-test (B,F,I) . RNA was also isolated from 10T1/2, LR21, RHAMM Δexon8-16 , KOV4, and KOV5 cell lines, with mRNA levels for mTert , Pinx1, Sirt1, Tpp1 , and Pot1a measured similarly (C,D,E,G,J) . For RHAMM Δexon8-16 (KO) and rescued lines (KOV4 and KOV5), treatment with PD98059 at 50 µM for 24 h was followed by qRT-PCR analysis to measure the mRNA expression of Tpp1 , and Pot1a , normalized to Gapdh . Results are means ± SD; n = 2 experiments; *p < 0.05, **p < 0.01, ***p < 0.001 (H,K) .

Article Snippet: The TRAPeze Gel-based Telomerase Detection Kit (Cat #S7700, Millipore Sigma, United States) was used to perform the telomerase repeat amplification protocol (TRAP) assay according to the manufacturer’s instructions.

Techniques: CRISPR, Isolation, Quantitative RT-PCR, Expressing

Journal: Frontiers in Aging

Article Title: The truncated isoform of the receptor for hyaluronan-mediated motility (RHAMM Δ163 ) modulates shelterin and telomerase reverse transcriptase transcription affecting telomerase activity

doi: 10.3389/fragi.2025.1604051

Figure Lengend Snippet: Telomerase activity affects RHAMM expression. Cycloastragenol (3 µM) was added to 10T1/2 fibroblasts for 6 h, followed by RNA isolation and qRT-PCR analysis to measure the mRNA expression of Hmmr, mTert, Tpx2 , Aurka , and Has2, normalized to Gapdh or Snrpd3 . Data are means ± SD; n = 3 experiments; *p < 0.05, **p < 0.01, ***p < 0.001 by Student’s t-test (A–E) . The extracellular hyaluronan concentration was analyzed by hyaluronan-based ELISA assay. The hyaluronan level was normalized by total RNA concentration (F) .

Article Snippet: The TRAPeze Gel-based Telomerase Detection Kit (Cat #S7700, Millipore Sigma, United States) was used to perform the telomerase repeat amplification protocol (TRAP) assay according to the manufacturer’s instructions.

Techniques: Activity Assay, Expressing, Isolation, Quantitative RT-PCR, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Loss of function of Atrx recapitulates phenotypes of alternative lengthening of telomeres in a primary mouse model of sarcoma

doi: 10.1016/j.isci.2025.112357

Figure Lengend Snippet: Characterizing telomere length and telomerase function (A) Telomere restriction fragment assay of RsaI and HinfI restricted DNA from tumors “T” and normal liver controls “L” visualized using chemiluminescent detection. (B) Telomerase Repeated Amplification Protocol (TRAP) assay performed on cell lines derived from primary tumors. 143B is an osteosarcoma cell line with known telomerase activity (positive control), and U2OS is an osteosarcoma cell line without functional telomerase (negative control).

Article Snippet: TRAPeze® Telomerase Detection Kit , Millipore Sigma , S7700.

Techniques: Amplification, TRAP Assay, Derivative Assay, Activity Assay, Positive Control, Functional Assay, Negative Control

Journal: iScience

Article Title: Loss of function of Atrx recapitulates phenotypes of alternative lengthening of telomeres in a primary mouse model of sarcoma

doi: 10.1016/j.isci.2025.112357

Figure Lengend Snippet:

Article Snippet: TRAPeze® Telomerase Detection Kit , Millipore Sigma , S7700.

Techniques: Recombinant, Blocking Assay, Staining, Bicinchoninic Acid Protein Assay, Software