human fetal lung fibroblast cell line  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: HDAC Class I Inhibitor Domatinostat Induces Apoptosis Preferentially in Glioma Stem Cells Through p53-Dependent and -Independent Activation of BAX Expression

doi: 10.3390/ijms26167803

Figure Lengend Snippet: Domatinostat preferentially inhibits cell growth and induces cell death in glioma stem cells. ( A ) GS-Y01, GS-Y03, TGS01, their differentiated counterparts (dGS-Y01, dGS-Y03, and dTGS01), and IMR-90 human lung fibroblasts were treated with the indicated concentrations of domatinostat for 1–4 days, and the number of viable cells was assessed by trypan blue dye exclusion. ( B ) Cells were treated as in ( A ), and the percentage of dead cells was evaluated using the trypan blue dye exclusion assay. * p < 0.05 vs. treatment without domatinostat (i.e., at 0 nM).

Article Snippet: IMR-90, a human normal fetal lung fibroblast cell line, was purchased from the American Type Culture Collection (Manassas, VA, USA) and maintained in DMEM supplemented with 10% FBS.

Techniques: Exclusion Assay

Journal: International Journal of Molecular Sciences

Article Title: HDAC Class I Inhibitor Domatinostat Induces Apoptosis Preferentially in Glioma Stem Cells Through p53-Dependent and -Independent Activation of BAX Expression

doi: 10.3390/ijms26167803

Figure Lengend Snippet: Domatinostat induces apoptotic cell death. ( A ) GS-Y01, GS-Y03, TGS01, their differentiated counterparts (dGS-Y01, dGS-Y03, and dTGS01), and IMR-90 human lung fibroblasts were treated without or with 500 nM domatinostat for 1–3 days, and were then subjected to Western blot analyses of the indicated proteins. The numbers below the images represent the means ( n = 2) of the relative band intensities after each band was quantified by densitometry and normalized to the GAPDH value. * p < 0.05 vs. cells before domatinostat treatment (i.e., cells at 0 days) for each of the glioma stem cell lines and their differentiated counterparts, except for the right most panel where comparisons were made with the adjacent left lane (i.e., IMR-90 treated with domatinostat for 3 days). ( B ) Cells were treated without or with 500 nM domatinostat in the absence or presence of 40 μM Z-VAD-fmk for 1 day, and were then subjected to Western blot analyses of the indicated proteins. * p < 0.05 vs. cells treated with domatinostat alone. ( C ) Cells were treated as in ( B ), and were then subjected to the propidium iodide (PI) incorporation assay to assess the percentage of dead cells. * p < 0.05. In ( A , B ), the expected migrating position for cleaved caspase-3 is indicated by an arrow.

Article Snippet: IMR-90, a human normal fetal lung fibroblast cell line, was purchased from the American Type Culture Collection (Manassas, VA, USA) and maintained in DMEM supplemented with 10% FBS.

Techniques: Western Blot

Journal: bioRxiv

Article Title: Targeted intervention of senescence induced by a 5′ half fragment of tRNA Seca(NCA) with antisense oligonucleotide extends healthspan and lifespan in mice

doi: 10.1101/2025.03.24.644848

Figure Lengend Snippet: a , CCK-8 assay showed that the proliferation of TCMK-1 and 2BS treated with 5′-tRNA Sec(NCA) half was significantly inhibited compared with the blank transfection reagent group. b . SA-β-gal staining indicated that the SA-β-gal positive rate of TCMK-1 and 2BS cells treated with 5′-tRNA Sec(NCA) half increased significantly, presenting a cellular senescence phenotype. c , The expression levels of senescence markers P21, P16, IL6, and CCL2 were significantly upregulated in cells induced by 5′-tRNA Sec(NCA) half. d , ROS significantly increased in cells treated with 5′-tRNA Sec(NCA) half. e , The telomere length (T/S ratio) of cells treated with 5′-tRNA Sec(NCA) half significantly decreased. f . IFN-β levels significantly upregulated in cells treated with 5′-tRNA Sec(NCA) half, while the level of IFN-γ did not exhibit such huge change. g-h , Liposome nanoparticle (LNP) delivery system encapsulated with 5′-tRNA Sec(NCA) half was intravenously injected into young Balb/c female mice. After 6 days, the senescence markers in the kidneys of mice were significantly upregulated. Data presented in c , d , e , and f are presented as mean ± S.D. Data presented in h are presented as mean ± S.E.M. Statistical test: two-tailed unpaired t -test. * P <0.05; ** P <0.01; *** P <0.001; **** P <0.0001; n.s., non-significant.

Article Snippet: 2BS human fetal lung fibroblasts cell line purchased from American Type Culture Collection was cultured in Minimum Essential Medium (MEM) supplemented with 10% fetal bovine serum (FBS), 1% non-essential amino acids (NEAA), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: CCK-8 Assay, Transfection, Staining, Expressing, Injection, Two Tailed Test

Journal: bioRxiv

Article Title: Targeted intervention of senescence induced by a 5′ half fragment of tRNA Seca(NCA) with antisense oligonucleotide extends healthspan and lifespan in mice

doi: 10.1101/2025.03.24.644848

Figure Lengend Snippet: a , Transcriptomic sequencing revealed that dsRNA sensor genes significantly upregulated in cells treated with 5′-tRNA Sec(NCA) half. b , qPCR analysis confirmed that the expression of the above genes and MYD88, a key adaptor protein in the TLR7 signaling pathway, increased significantly. c , In TCMK-1 and 2BS cells treated with 5′-tRNA Sec(NCA) half, the protein level of TLR7 was significantly upregulated in a dose-dependent manner. d , Simulation of the interaction between 5′-tRNA Sec(NCA) half and TLR7 protein predicted by AlphaFold 3. e , RNA pull-down assay demonstrated that 5′-tRNA Sec(NCA) half specifically bound to TLR7. f-g , In TLR7-knockdown (TLR7-KD) cells, co-treatment with 5′-tRNA Sec(NCA) half and siTLR7 led to a significant downregulation of senescence markers and a lower SA-β-gal positive rate compared with treatment with 5′-tRNA Sec(NCA) half alone. Data presented in b , c , and f are presented as mean ± S.D. Statistical test: two-tailed unpaired t -test. * P <0.05; ** P <0.01; *** P <0.001; **** P <0.0001.

Article Snippet: 2BS human fetal lung fibroblasts cell line purchased from American Type Culture Collection was cultured in Minimum Essential Medium (MEM) supplemented with 10% fetal bovine serum (FBS), 1% non-essential amino acids (NEAA), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Sequencing, Expressing, Pull Down Assay, Knockdown, Two Tailed Test

Journal: bioRxiv

Article Title: Targeted intervention of senescence induced by a 5′ half fragment of tRNA Seca(NCA) with antisense oligonucleotide extends healthspan and lifespan in mice

doi: 10.1101/2025.03.24.644848

Figure Lengend Snippet: a , 5′-tRNA Sec(NCA) half significantly increased in D-galactose-induced TCMK-1 aged cells and 2BS naturally aged cells. b , CCK-8 assay indicated that the proliferation of aged cells treated with ASO-5′-tRNA Sec(NCA) half significantly increased compared with the liposome-treated controls. c . Treatment with ASO-5′-tRNA Sec(NCA) half resulted in a significant decrease in the number of SA-β-gal-positive cells in aged cells. d , ROS levels were markedly reduced in aged cells treated with ASO-5′-tRNA Sec(NCA) half. e-f . ASO-5′-tRNA Sec(NCA) half significantly downregulated aging markers and extended telomere length in aged cells. g , LNP delivery of ASO-5′-tRNA Sec(NCA) half significantly extended the lifespan of paraquat-induced acute aged mice in a dose-dependent manner, as well as dysregulating TLR7, P21 and IL6 in multiple organs ( h ). Data presented in a , b , d , e , f and h are presented as mean ± S.D. Data presented in g are presented as log-rank test and the Gehan-Breslow-Wilcoxon. Statistical test: two-tailed unpaired t -test. * P <0.05; ** P <0.01; *** P <0.001; **** P <0.0001; n.s., non-significant.

Article Snippet: 2BS human fetal lung fibroblasts cell line purchased from American Type Culture Collection was cultured in Minimum Essential Medium (MEM) supplemented with 10% fetal bovine serum (FBS), 1% non-essential amino acids (NEAA), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: CCK-8 Assay, Two Tailed Test