Journal: bioRxiv
Article Title: STING safeguards epithelial genome integrity and protects from carcinogenesis via mitotic checkpoint control
doi: 10.64898/2026.02.17.706442
Figure Lengend Snippet: a : Western blot analysis of γH2AX from MODE-K WT and STING -/- cell lines after treated with 2.5μM AraC for 20h. b-c : Representative picture (b) and statistical analysis of tail length (c) measured from comet assay from MODE-K WT and STING -/- cell lines after treated with 2.5μM AraC for 20h. Significance was determined using unpaired two-tailed student’s test. d : Western blot analysis of ATM from MODE-K WT and STING -/- cell lines after treated with 2.5μM AraC for 20h. e-f : Scheme of DR-GFP reporter assay ( e ). HR levels of MODE-K WT and STING -/- cell lines were measured by FACS ( f ). Significance was determined using unpaired two-tailed student’s test. g : MODE-K WT and STING -/- cells were transfected with either siRNA against Wip1 or control siRNA for 24h then stimulted with 2.5μM AraC for 20h. Protein levels were measured by western blot assay. h : Co-immunoprecipitation (co-IP) analysis of the interaction between ATM and NBS1 from MODE-K WT and STING-/- cell lines after treated with 2.5μM AraC for 20h. ATM-containing complexes were immunoprecipitated using an anti-ATM antibody and probed for NBS1 by immunoblotting. i : Scheme of protein kinase assay. WT and STING cell lines were treated with 2.5 μM AraC for 20 hours. Following protein extraction, serine/threonine kinase activity and tyrosine kinase activities were analyzed using the Pamgene kinase assay. j : Box plots showing the sum of log2-transformed fluorescence signal intensities of serine/threonine-kinase (STK) phosphorylation levels on the chip. k : Deeper look into different kinase families that are affected by STING deficiency, full figure shown in . l : Deeper look into the results from upstream kinase analysis showing the main affected STK, the full figure shown in . m : MODE-K WT, STING -/- , and cGAS -/- cells were treated with 2.5 μM AraC for 20h. Protein levels were analyzed by western blot. For all the significance analysis: ns = not significant, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.
Article Snippet: Cells were transiently transfected with the DR-GFP reporter plasmid (Addgene, #26475) using Lipofectamine TM 3000 Reagent (Thermofisher, L3000015) according to the manufacturer’s instructions.
Techniques: Western Blot, Single Cell Gel Electrophoresis, Two Tailed Test, Reporter Assay, Transfection, Control, Immunoprecipitation, Co-Immunoprecipitation Assay, Protein Kinase Assay, Protein Extraction, Activity Assay, Kinase Assay, Transformation Assay, Fluorescence, Phospho-proteomics
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