human fetal lung fibroblast wi 38  (ATCC)

Journal: Molecular Therapy Oncology

Article Title: Tackling cancer heterogeneity with systemically delivered oncolytic adenoviruses transcriptionally targeted with hybrid promoters

doi: 10.1016/j.omton.2025.201073

Figure Lengend Snippet: AR2015 lytic activity in vitro compared to single TSP-driven OAds (A–I) In vitro lytic activity of the OAds AR2015, AV22EL, and AV636, as well as Ad5WT (positive control), in CRC cell lines LoVo, T84, HT29, and HCT116, normal human colonic epithelial cells (CCD841), human fetal lung fibroblasts (WI-38), human microendothelial cells (HMEC-1), and human melanoma cells (A375 and SB2). Cells (1 × 10 4 ) were seeded in 24-well plates and infected 24 h later with increasing multiplicities of infection (MOIs: 0–100). After 6 days, cell viability was assessed using the MTS assay and expressed as mean ± SD ( n = 3), with the viability of uninfected control cells set to 100%. Two-way ANOVA was performed for statistical analysis, followed by Dunnett’s test (vs. Ad(F5)WT). (J) Comparative replication kinetics of AR2015 and AV22EL in CRC cell lines (LoVo, T84, HT29, and HCT116) and normal CCD841 cells. Cells were infected at an MOI of 100, and samples were collected at 5 h (baseline) and 72 h post-infection. Viral replication was quantified by qPCR targeting the adenoviral E4 gene. Data are presented as a fold increase in E4 levels (72 vs. 5 h). (K) Expression of adenoviral E1A protein in LoVo cells at 8 and 24 h post-infection with AV22EL, AR2015, or Ad5WT. Protein levels were assessed by western blot, with β-actin used as a loading control.

Article Snippet: Human CRC cell lines (LoVo, T84, HCT116, and HT29), normal colon epithelial cells (CCD841), human embryonic kidney cells (HEK293), human fetal lung fibroblasts (WI-38), and human microendothelial cells (HMEC-1) were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Activity Assay, In Vitro, Positive Control, Infection, MTS Assay, Control, Expressing, Western Blot

Journal: Aging cell

Article Title: hTERT Increases TRF2 to Induce Telomere Compaction and Extend Cell Replicative Lifespan.

doi: 10.1111/acel.70105

Figure Lengend Snippet: FIGURE 3 | hTERT variants regulate the TRF2:TRF1 ratio by transcriptional and posttranslational mechanisms. (A) TRF2 mRNA levels deter- mined by RT-PCR in young and senescent BJ, Hs68, and Wi38 fibroblast strains. (B) Western blots of young (Y) and senescent (S) fibroblast strains probed with the indicated antibodies. (C) TRF1 mRNA levels in young and senescent BJ cells and BJs expressing WT-hTERT, R865C-hTERT, and V144M-hTERT variants. (D) Western blots of young and senescent BJ fibroblasts, and BJ cells expressing hTERT variants, probed with the indicated antibodies. (E) TRF2 protein levels in young and senescent fibroblast strains. (F) TRF2 protein levels in young and senescent BJs and BJs expressing hTERT variants. (A,C,E,F) Error bars = 95% CI; n = 3. p values determined by two-tailed unpaired t-tests with Welch's correction, ns = not significant. (G) Quantitation of the levels of the CDC20 and (H) FBXO5 Siah1 e3 ubiquitin ligases in cells expressing hTERT variants. Graphs show three bio- logical replicates with tables showing densitometry values for each replicate, with young BJ fibroblasts set to a value of 1. Error bars = 95% CI; n = 3. p values determined by two-tailed unpaired t-tests with Welch's correction. (I) Mock-transfected cells or cells expressing Siah1, CDC20, or FBXO5 were harvested 48 h after transfection, and lysates were blotted for the indicated proteins. GAPDH served as the loading control.

Article Snippet: Primary Hs68 human foreskin fibroblasts (ATCC CRL- 1635) and WI38 fetal lung fibroblasts (ATCC CCL- 75) were grown in DMEM, 1 g/L glucose (Gibco, 11885- 084), supplemented with 10% (v/v) FBS.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Two Tailed Test, Quantitation Assay, Ubiquitin Proteomics, Transfection, Control