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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: Rabbit polyclonal anti-53BP1 , Novus Biologicals , Cat#NB 100-304, RRID: AB_350221.
Techniques: Control, Expressing