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: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Characterization of human breast cancer epithelial cells (HBCEC) derived from long term cultured biopsies

doi: 10.1186/1756-9966-28-127

Figure Lengend Snippet: Surface marker expression, SA-β-gal staining and telomerase activity in HBCEC . A . Determination of the percentage of cell surface marker expression in HBCEC at different ages. Expression of the surface marker proteins CD24, CD44, CD227 was maintained during long term culture of HBCEC. Whereas CD24 and CD44 were similarly expressed after 176d and 462d, CD227 increased from 52% to 88% in HBCEC 462d. The flow cytometry measurements varied by about 8%. B . SA-β-gal staining of primary HBCEC and HMEC cultures. Staining for SA-β-gal of a HBCEC population after 722d in culture revealed little if any positive cell. Normal HMEC in passage 16, however, displayed already predominantly enlarged senescent cells after 32d, demonstrated by the dark-green stain (bar = 200 μm). C . Telomerase (TRAP-)assay of primary cultures from breast cancer biopsies. Telomerase activity was analyzed according to the Telomeric Repeat Amplification Protocol (TRAP). HBCEC populations demonstrated telomerase activity independent of the age of the culture and the harvest method. The human embryonic kidney (HEK) 293T cell line was used as a positive control and 1× CHAPS buffer served as a negative control. Quantification was performed using densitometric analysis.

Article Snippet: The TRAPEZE ® Gel-Based Telomerase Detection assay (Chemicon International, Temecula, CA, USA) was performed according to the manufacturer's protocol using the isotopic detection.

Techniques: Marker, Expressing, Staining, Activity Assay, Flow Cytometry, TRAP Assay, Amplification, Positive Control, Negative Control