Journal: iScience

Article Title: The effect of the probiotic consortia on SARS-CoV-2 infection in ferrets and on human immune cell response in vitro

doi: 10.1016/j.isci.2022.104445

Figure Lengend Snippet:

Article Snippet: Streptococcus thermophilus subsp. thermophilus St-21 , ATCC , SD5207.

Techniques: Virus, Recombinant, Reverse Transcription, Isolation, Gene Expression, Software, Maltodextrin

Journal: iScience

Article Title: Human SKI component SKIV2L regulates telomeric DNA-RNA hybrids and prevents telomere fragility

doi: 10.1016/j.isci.2024.111096

Figure Lengend Snippet: SKIV2L and TTC37 prevents telomere fragility (A and B) Telomere FISH analysis in SKIV2L- and TTC37-depleted HeLa1.3 and HT1080-ST cells using siRNAs: % of telomere fragility (yellow) and loss (purple) per metaphase in siCtr, siSKIV2L and siTTC37 (means ± SD, n > 25 metaphases, 2 independent experiments, scale bar, 10 μm). t test ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001. (C) Telomere FISH analysis in shCtr (Control) or shSKIV2L HEK293 cells expressing GFP or SKIV2L (means ± SD, n > 25 metaphases, 2 independent experiments, scale bar, 10 μm). Details are as in (A). (D) Telomere FISH analysis in TTC37-depleted HEK293 cells using siRNAs (means ± SD, n = 25 metaphases). Details are as in (A). (E) Telomere FISH analysis in HeLa1.3 cells using shRNAs: shCtr and shSKIV2L treated with DMSO (−, control) or APH (+) ( n > 45 metaphases, 2 independent experiments). Details are as in (A). (F) IF of pre-extracted cells showing co-localization of pS1981 ATM autophosphorylation with TRF2 (telomeres) in shCtr and SKIV2L-depleted (shSKIV2L) HeLa1.3 cells. Quantification of the percentage of cells with co-localization foci and mean number of foci per nucleus is depicted (means ± SEM, n = 200 cells, 2 independent experiments, scale bar 15 μm). t test ∗∗∗ p < 0.001. (G) IF-FISH of pre-extracted cells showing co-localization of 53BP1 with telomeres in asynchronous (AS) or G2-synchronized control (shCtr) and SKIV2L-depleted (shSKIV2L) HeLa1.3 cells. Quantification of the percentage of cells with co-localization foci is depicted (means ± SEM, n > 400 cells, 3 independent experiments, scale bar 15 μm). t test ∗ p < 0.05. See also Figure S2 .

Article Snippet: Stable clone of adult female human cervical adenocarcinoma HeLa cell line with long telomeres (HeLa1.3 cells); adult male human fibrosarcoma HT1080 cell line overexpressing telomase (HT1080 super-telomerase cells, HT1080-ST) (kindly provided by T. de Lange and J. Lingner, respectively); adult female human osteosarcoma U2OS cell line (ATCC); human embryonic kidney HEK293FT cell line (ATCC); and human female fetal lung fibroblast IMR90 cell line (Coriel Institute) were cultured in DMEM medium supplemented with 10% (v/v) fetal bovine serum (FBS, Sigma-Aldrich, F2442) and maintained at 37°C in 5% (v/v) CO 2 .

Techniques: Control, Expressing

Journal: iScience

Article Title: Human SKI component SKIV2L regulates telomeric DNA-RNA hybrids and prevents telomere fragility

doi: 10.1016/j.isci.2024.111096

Figure Lengend Snippet: SKIV2L regulates telomeric DNA-RNA hybrids in cellulo to prevent telomere fragility (A) Proximity ligation assay (PLA) showing co-localization of SKIV2L and TRF2 in asynchronous (AS) and in G2-synchronized HeLa1.3 cells with and without RNase H1 (RNH1) overexpression (median, Q1 and Q3, at least 600 cells scored per condition, 4 independent experiments, scale bar 10 μm). Mann-Whitney U test ∗∗∗∗ p < 0.0001. (B) S9.6 IF in HeLa1.3 cells treated with RNAse III, with and without RNase H1 (RNH1) overexpression (median ± interquartile range, 360 cells scored per condition, 4 independent experiments, scale bar, 10 μm). Mann-Whitney U test ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (C) PLA showing the co-localization of DNA-RNA hybrids (S9.6) and TRF2 in HeLa1.3 cells pre-extracted and treated with RNAse III (median, Q1 and Q3, at least 400 cells scored per condition, 2 independent experiments, scale bar 10 μm). Mann-Whitney U test ∗∗∗∗ p < 0.0001. (D) DRIP showing the levels of DNA-RNA hybrids at telomeres in AS and G2-synchronized HeLa1.3 cells, RNH, RNase H treatment (means ± SEM, n = 4). (E) DRIP-qPCR assay of G2-synchronized HEK293 cells overexpressing GFP or SKIV2L at 10q, 13q, 20q, and 22q subtelomeric regions. RNH, RNase H treatment (means ± SEM, n = 5). Percent input values were normalized to the GFP overexpressing condition. (F) Model proposing the function of hSKI at telomeres. Telomeric DNA-RNA hybrid accumulation in late S/G2 phase drives hSKI recruitment to telomeres to regulate physiological DNA-RNA hybrid levels, prevent telomere replication stress and ensure telomere stability. See also Figure S5 .

Article Snippet: Stable clone of adult female human cervical adenocarcinoma HeLa cell line with long telomeres (HeLa1.3 cells); adult male human fibrosarcoma HT1080 cell line overexpressing telomase (HT1080 super-telomerase cells, HT1080-ST) (kindly provided by T. de Lange and J. Lingner, respectively); adult female human osteosarcoma U2OS cell line (ATCC); human embryonic kidney HEK293FT cell line (ATCC); and human female fetal lung fibroblast IMR90 cell line (Coriel Institute) were cultured in DMEM medium supplemented with 10% (v/v) fetal bovine serum (FBS, Sigma-Aldrich, F2442) and maintained at 37°C in 5% (v/v) CO 2 .

Techniques: Proximity Ligation Assay, Over Expression, MANN-WHITNEY

Journal: iScience

Article Title: Human SKI component SKIV2L regulates telomeric DNA-RNA hybrids and prevents telomere fragility

doi: 10.1016/j.isci.2024.111096

Figure Lengend Snippet:

Article Snippet: Stable clone of adult female human cervical adenocarcinoma HeLa cell line with long telomeres (HeLa1.3 cells); adult male human fibrosarcoma HT1080 cell line overexpressing telomase (HT1080 super-telomerase cells, HT1080-ST) (kindly provided by T. de Lange and J. Lingner, respectively); adult female human osteosarcoma U2OS cell line (ATCC); human embryonic kidney HEK293FT cell line (ATCC); and human female fetal lung fibroblast IMR90 cell line (Coriel Institute) were cultured in DMEM medium supplemented with 10% (v/v) fetal bovine serum (FBS, Sigma-Aldrich, F2442) and maintained at 37°C in 5% (v/v) CO 2 .

Techniques: Virus, Recombinant, Protease Inhibitor, Reverse Transcription, Blocking Assay, Mass Spectrometry, SYBR Green Assay, Flow Cytometry, Imaging, Mutagenesis, Cell Cycle Assay, shRNA, Software