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dr gfp reporter plasmid  (Addgene inc)


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    Addgene inc dr gfp reporter plasmid
    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 <t>of</t> <t>DR-GFP</t> 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 <t>were</t> <t>transfected</t> 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.
    Dr Gfp Reporter Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 188 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dr+gfp+reporter+plasmid/pDRGFP+(Plasmid+%2326475)/bio_rxiv__64898__2026__02__17__706442-227-6-9
    Average 96 stars, based on 188 article reviews
    dr gfp reporter plasmid - by Bioz Stars, 2026-10
    96/100 stars

    Images

    1) Product Images from "STING safeguards epithelial genome integrity and protects from carcinogenesis via mitotic checkpoint control"

    Article Title: STING safeguards epithelial genome integrity and protects from carcinogenesis via mitotic checkpoint control

    Journal: bioRxiv

    doi: 10.64898/2026.02.17.706442

    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.
    Figure Legend 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.

    Techniques Used: 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

    Related Articles

    Plasmid Preparation:

    Article Title: SMARCAD1-mediated active replication fork stability maintains genome integrity
    Article Snippet: .. After being seeded in a six-well plate overnight, cells were cotransfected with DR-GFP reporter plasmid (Addgene #26475) and I-Scel expression vector (Addgene #26477) or empty vector using X-tremeGENE 9 DNA transfection agent (Roche) according to the manufacturer’s protocol for two consecutive days. p-MAX-GFP plasmid (Addgene #16007) was transfected in parallel to assess transfection efficiency. ..

    Article Title: STING safeguards epithelial genome integrity and protects from carcinogenesis via mitotic checkpoint control
    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. .. After 24 h, cells were re-transfected with DR-GFP together with the I-SceI expression plasmid (Addgene, #26477) to induce site-specific DSBs.

    Article Title: ATR-mediated proteome remodeling is a major determinant of homologous recombination capacity in cancer cells
    Article Snippet: Primary human fibroblast cells from control subjects (GM08398) and patients with Seckel syndrome (GM18366, ATR-defective) were obtained from Coriell Institute for Medical Research, Camden, NJ, USA. .. For the DR-GFP assay, cells (U2OS, HCT116, hTERT RPE-1, HEK293T, HeLa, U87, T98G, A2780, 293-T-REX-E2F6 cells and SV40 large T antigen transformed hTERT RPE-1) were transfected with the mCherry plasmid or DR-GFP reporter plasmid (pDR-GFP; addgene plasmid 26475) together with a plasmid coding for I-SceI (pCBASceI; addgene plasmid 26477; gifts from M. Jasin) or DR-GFP reporter plasmid together with an empty plasmid pCAGGS. .. Two days after transfection, cells were trypsinized, resuspended in PBS and then analyzed by flow cytometry using FACSAria Fusion or Accuri C6 cytometer (BD).

    Article Title: SMARCAD1 Mediated Active Replication Fork Stability Maintains Genome Integrity
    Article Snippet: .. After being seeded in a 6-well plate for 24 hours, cells were co-transfected with 1.5 μg of DR-GFP reporter plasmid (addgene #26475) and 1.5 μg I-Scel expression vector (addgene # 26477) or empty vector using X-tremeGENE 9 DNA transfection agent (Roche) according to the manufacturer’s protocol for 24 hours. p-MAX-GFP plasmid (addgene #16007) was transfected in parallel to assess transfection efficiency. ..

    Expressing:

    Article Title: SMARCAD1-mediated active replication fork stability maintains genome integrity
    Article Snippet: .. After being seeded in a six-well plate overnight, cells were cotransfected with DR-GFP reporter plasmid (Addgene #26475) and I-Scel expression vector (Addgene #26477) or empty vector using X-tremeGENE 9 DNA transfection agent (Roche) according to the manufacturer’s protocol for two consecutive days. p-MAX-GFP plasmid (Addgene #16007) was transfected in parallel to assess transfection efficiency. ..

    Article Title: SMARCAD1 Mediated Active Replication Fork Stability Maintains Genome Integrity
    Article Snippet: .. After being seeded in a 6-well plate for 24 hours, cells were co-transfected with 1.5 μg of DR-GFP reporter plasmid (addgene #26475) and 1.5 μg I-Scel expression vector (addgene # 26477) or empty vector using X-tremeGENE 9 DNA transfection agent (Roche) according to the manufacturer’s protocol for 24 hours. p-MAX-GFP plasmid (addgene #16007) was transfected in parallel to assess transfection efficiency. ..

    Transfection:

    Article Title: SMARCAD1-mediated active replication fork stability maintains genome integrity
    Article Snippet: .. After being seeded in a six-well plate overnight, cells were cotransfected with DR-GFP reporter plasmid (Addgene #26475) and I-Scel expression vector (Addgene #26477) or empty vector using X-tremeGENE 9 DNA transfection agent (Roche) according to the manufacturer’s protocol for two consecutive days. p-MAX-GFP plasmid (Addgene #16007) was transfected in parallel to assess transfection efficiency. ..

    Article Title: STING safeguards epithelial genome integrity and protects from carcinogenesis via mitotic checkpoint control
    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. .. After 24 h, cells were re-transfected with DR-GFP together with the I-SceI expression plasmid (Addgene, #26477) to induce site-specific DSBs.

    Article Title: ATR-mediated proteome remodeling is a major determinant of homologous recombination capacity in cancer cells
    Article Snippet: Primary human fibroblast cells from control subjects (GM08398) and patients with Seckel syndrome (GM18366, ATR-defective) were obtained from Coriell Institute for Medical Research, Camden, NJ, USA. .. For the DR-GFP assay, cells (U2OS, HCT116, hTERT RPE-1, HEK293T, HeLa, U87, T98G, A2780, 293-T-REX-E2F6 cells and SV40 large T antigen transformed hTERT RPE-1) were transfected with the mCherry plasmid or DR-GFP reporter plasmid (pDR-GFP; addgene plasmid 26475) together with a plasmid coding for I-SceI (pCBASceI; addgene plasmid 26477; gifts from M. Jasin) or DR-GFP reporter plasmid together with an empty plasmid pCAGGS. .. Two days after transfection, cells were trypsinized, resuspended in PBS and then analyzed by flow cytometry using FACSAria Fusion or Accuri C6 cytometer (BD).

    Article Title: SMARCAD1 Mediated Active Replication Fork Stability Maintains Genome Integrity
    Article Snippet: .. After being seeded in a 6-well plate for 24 hours, cells were co-transfected with 1.5 μg of DR-GFP reporter plasmid (addgene #26475) and 1.5 μg I-Scel expression vector (addgene # 26477) or empty vector using X-tremeGENE 9 DNA transfection agent (Roche) according to the manufacturer’s protocol for 24 hours. p-MAX-GFP plasmid (addgene #16007) was transfected in parallel to assess transfection efficiency. ..

    Transformation Assay:

    Article Title: ATR-mediated proteome remodeling is a major determinant of homologous recombination capacity in cancer cells
    Article Snippet: Primary human fibroblast cells from control subjects (GM08398) and patients with Seckel syndrome (GM18366, ATR-defective) were obtained from Coriell Institute for Medical Research, Camden, NJ, USA. .. For the DR-GFP assay, cells (U2OS, HCT116, hTERT RPE-1, HEK293T, HeLa, U87, T98G, A2780, 293-T-REX-E2F6 cells and SV40 large T antigen transformed hTERT RPE-1) were transfected with the mCherry plasmid or DR-GFP reporter plasmid (pDR-GFP; addgene plasmid 26475) together with a plasmid coding for I-SceI (pCBASceI; addgene plasmid 26477; gifts from M. Jasin) or DR-GFP reporter plasmid together with an empty plasmid pCAGGS. .. Two days after transfection, cells were trypsinized, resuspended in PBS and then analyzed by flow cytometry using FACSAria Fusion or Accuri C6 cytometer (BD).



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    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 <t>of</t> <t>DR-GFP</t> 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 <t>were</t> <t>transfected</t> 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.
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    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 <t>of</t> <t>DR-GFP</t> 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 <t>were</t> <t>transfected</t> 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.
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    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 <t>of</t> <t>DR-GFP</t> 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 <t>were</t> <t>transfected</t> 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.
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    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 <t>of</t> <t>DR-GFP</t> 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 <t>were</t> <t>transfected</t> 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.
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    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 <t>of</t> <t>DR-GFP</t> 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 <t>were</t> <t>transfected</t> 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.
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    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.

    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