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Addgene inc dr jen liou
Dr Jen Liou, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 20 article reviews

dr jen liou - by Bioz Stars, 2026-05

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(A) Survival of HeLa RAD54B KO cells transfected with the siCTRL, siRAD54L or siBRCA2 upon treatment with 5 µM of olaparib or untreated. Cell viability was measured using growth curve analyses. All data show mean ± SEM (n=3). (B) HeLa (i) EV, (ii) RAD54L KO, (iii) RAD51AP1 KO and (iv) RAD54B KO cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated after treatment with 1 µM olaparib for 24 h in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 2-3). Results from individual experiments, each derived from 40 cells, are indicated. (C) HeLa pGC reporter cells carry a GFP cassette that is disrupted by the I-SceI sequence and a second truncated GFP cassette (left). I-SceI-induced HR repair events result in the expression of functional GFP. HeLa pGC cells were transfected with the indicated siRNAs, followed by transfection with the I-SceI expression plasmid to induce site-specific DSBs. The percentage of GFP-positive cells was measured by flow cytometry. All data show mean ± SEM (n=3). All data show mean ± SEM (n = 3). Results from individual experiments are indicated. (D) (Top) U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated after treatment with 1 µM olaparib for 24 h in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 2-5). Results from individual experiments, each derived from 40 cells, are indicated. (Bottom) Knockdown of BRCA2, RAD51AP1 and RAD54L was confirmed by western blotting. (E) Quantitative RT–PCR analysis of RAD54B mRNA expression levels in HeLa and U2OS cells after transfection with siCTRL or siRAD54B. Expression levels were normalized to GAPDH, with siCTRL defined as 100%. All data show mean ± SEM (n = 2). (F) U2OS DR-GFP cells were transfected with the indicated siRNAs, followed by transfection with the I-SceI expression plasmid to induce site-specific DSBs. The percentage of GFP-positive cells, representing HR repair events, was measured by flow cytometry. All data show mean ± SEM (n = 4). Results from individual experiments are indicated. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns: not significant (two-tailed t test).

Journal: bioRxiv

Article Title: PARP inhibitor synthetic lethality reveals homologous recombination sub-pathway architecture

doi: 10.64898/2026.03.06.709881

Figure Lengend Snippet: (A) Survival of HeLa RAD54B KO cells transfected with the siCTRL, siRAD54L or siBRCA2 upon treatment with 5 µM of olaparib or untreated. Cell viability was measured using growth curve analyses. All data show mean ± SEM (n=3). (B) HeLa (i) EV, (ii) RAD54L KO, (iii) RAD51AP1 KO and (iv) RAD54B KO cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated after treatment with 1 µM olaparib for 24 h in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 2-3). Results from individual experiments, each derived from 40 cells, are indicated. (C) HeLa pGC reporter cells carry a GFP cassette that is disrupted by the I-SceI sequence and a second truncated GFP cassette (left). I-SceI-induced HR repair events result in the expression of functional GFP. HeLa pGC cells were transfected with the indicated siRNAs, followed by transfection with the I-SceI expression plasmid to induce site-specific DSBs. The percentage of GFP-positive cells was measured by flow cytometry. All data show mean ± SEM (n=3). All data show mean ± SEM (n = 3). Results from individual experiments are indicated. (D) (Top) U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated after treatment with 1 µM olaparib for 24 h in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 2-5). Results from individual experiments, each derived from 40 cells, are indicated. (Bottom) Knockdown of BRCA2, RAD51AP1 and RAD54L was confirmed by western blotting. (E) Quantitative RT–PCR analysis of RAD54B mRNA expression levels in HeLa and U2OS cells after transfection with siCTRL or siRAD54B. Expression levels were normalized to GAPDH, with siCTRL defined as 100%. All data show mean ± SEM (n = 2). (F) U2OS DR-GFP cells were transfected with the indicated siRNAs, followed by transfection with the I-SceI expression plasmid to induce site-specific DSBs. The percentage of GFP-positive cells, representing HR repair events, was measured by flow cytometry. All data show mean ± SEM (n = 4). Results from individual experiments are indicated. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns: not significant (two-tailed t test).

Article Snippet: U2OS (ATCC), U2OS DR-GFP, HeLa-S3 cells (ATCC), HeLa EV, HeLa RAD54L, HeLa RAD54B KO, HeLa RAD51AP1 KO, HeLa ATRX KO and HeLa pGC cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Sigma), supplemented with 10% fetal bovine serum (FBS; Bio&SELL), 1% non-essential amino acid (NEAA; Sigma-Aldrich) and 1% penicillin/streptomycin (Sigma-Aldrich).

Techniques: Transfection, Derivative Assay, Sequencing, Expressing, Functional Assay, Plasmid Preparation, Flow Cytometry, Knockdown, Western Blot, Quantitative RT-PCR, Two Tailed Test

U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated 10 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns: not significant (two-tailed t test).

Journal: bioRxiv

Article Title: PARP inhibitor synthetic lethality reveals homologous recombination sub-pathway architecture

doi: 10.64898/2026.03.06.709881

Figure Lengend Snippet: U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated 10 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns: not significant (two-tailed t test).

Techniques: Transfection, Derivative Assay, Two Tailed Test

(A) Representative images of γH2AX foci and ATRX expression in U2OS ATRX G2 cells following transfection with the indicated siRNAs and subsequent irradiation are shown. (B) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs, and γH2AX foci were enumerated at 12 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown was confirmed by western blotting. (C) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs. They were irradiated with 2 Gy, collected after 16h, and processed to obtain chromosome spreads. SCEs per spread were quantified and normalized to 70 chromosomes. Results from individual experiments, each derived from 40 cells, are indicated (top), and red horizontal lines indicate the mean. The bar graph shows the IR-induced SCE numbers (total SCEs after background subtraction) (bottom). IR-induced SCE data show mean ± SEM, and results from individual experiments are indicated. 80-120 spreads and >7,000 chromosomes per condition were analysed from 2-3 independent experiments. (D) Model illustrating the interplay between RAD51AP1 and RAD54B, which promote SDSA, and RAD54L and ATRX, which facilitate dHJ formation.

Journal: bioRxiv

Article Title: PARP inhibitor synthetic lethality reveals homologous recombination sub-pathway architecture

doi: 10.64898/2026.03.06.709881

Figure Lengend Snippet: (A) Representative images of γH2AX foci and ATRX expression in U2OS ATRX G2 cells following transfection with the indicated siRNAs and subsequent irradiation are shown. (B) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs, and γH2AX foci were enumerated at 12 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown was confirmed by western blotting. (C) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs. They were irradiated with 2 Gy, collected after 16h, and processed to obtain chromosome spreads. SCEs per spread were quantified and normalized to 70 chromosomes. Results from individual experiments, each derived from 40 cells, are indicated (top), and red horizontal lines indicate the mean. The bar graph shows the IR-induced SCE numbers (total SCEs after background subtraction) (bottom). IR-induced SCE data show mean ± SEM, and results from individual experiments are indicated. 80-120 spreads and >7,000 chromosomes per condition were analysed from 2-3 independent experiments. (D) Model illustrating the interplay between RAD51AP1 and RAD54B, which promote SDSA, and RAD54L and ATRX, which facilitate dHJ formation.

Techniques: Expressing, Transfection, Irradiation, Derivative Assay, Knockdown, Western Blot

(A) (Top) U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated 10 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3-9). Results from individual experiments, each derived from 40 cells, are indicated. (Bottom) Knockdown of TOP3A, RECQ5, RAD51AP1 and RAD54L was confirmed by western blotting. (B) U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated after treatment with 1 µM olaparib for 24 h in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. (C) U2OS DR-GFP cells were transfected with the indicated siRNAs, followed by transfection with the I-SceI expression plasmid to induce site-specific DSBs. The percentage of GFP-positive cells, representing HR repair events, was measured by flow cytometry. All data show mean ± SEM (n = 3-7). Results from individual experiments are indicated. (D) U2OS cells were transfected with siCTRL, siBRCA2 and siTOP3A, irradiated, and RAD51 foci were enumerated 2 h following 2 Gy IR in EdU-negative G2 cells (left). Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown of BRCA2 and TOP3A was confirmed by western blotting (right). (E) U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated 10 h following 2 Gy IR in EdU-negative G2 cells (left). Spontaneous foci were subtracted. All data show mean ± SEM (n = 2-4). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown of BLM was confirmed by western blotting (right). *P < 0.05; **P < 0.01; ***P < 0.001; ns: not significant (two-tailed t test).

Journal: bioRxiv

Article Title: PARP inhibitor synthetic lethality reveals homologous recombination sub-pathway architecture

doi: 10.64898/2026.03.06.709881

Figure Lengend Snippet: (A) (Top) U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated 10 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3-9). Results from individual experiments, each derived from 40 cells, are indicated. (Bottom) Knockdown of TOP3A, RECQ5, RAD51AP1 and RAD54L was confirmed by western blotting. (B) U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated after treatment with 1 µM olaparib for 24 h in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. (C) U2OS DR-GFP cells were transfected with the indicated siRNAs, followed by transfection with the I-SceI expression plasmid to induce site-specific DSBs. The percentage of GFP-positive cells, representing HR repair events, was measured by flow cytometry. All data show mean ± SEM (n = 3-7). Results from individual experiments are indicated. (D) U2OS cells were transfected with siCTRL, siBRCA2 and siTOP3A, irradiated, and RAD51 foci were enumerated 2 h following 2 Gy IR in EdU-negative G2 cells (left). Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown of BRCA2 and TOP3A was confirmed by western blotting (right). (E) U2OS cells were transfected with the indicated siRNAs, and γH2AX foci were enumerated 10 h following 2 Gy IR in EdU-negative G2 cells (left). Spontaneous foci were subtracted. All data show mean ± SEM (n = 2-4). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown of BLM was confirmed by western blotting (right). *P < 0.05; **P < 0.01; ***P < 0.001; ns: not significant (two-tailed t test).

Techniques: Transfection, Derivative Assay, Knockdown, Western Blot, Expressing, Plasmid Preparation, Flow Cytometry, Irradiation, Two Tailed Test

(A) Representative images of γH2AX foci in U2OS ATRX G2 cells following transfection with the indicated siRNAs and subsequent irradiation are shown. (B) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs, and γH2AX foci were enumerated at 12 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. Striped bars indicate TOP3A depletion. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown was confirmed by western blotting. (C) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs. They were irradiated with 2 Gy, collected after 16h, and processed to obtain chromosome spreads. SCEs per spread were quantified and normalized to 70 chromosomes. Results from individual experiments, each derived from 40 cells, are indicated (left), and red horizontal lines indicate the mean. The bar graph shows the IR-induced SCE numbers (total SCEs after background subtraction) (right). IR-induced SCE data show mean ± SEM, and results from individual experiments are indicated. 120 spreads and >7,000 chromosomes per condition were analysed from three independent experiments. (D) U2OS cells were transfected with siCTRL, siRAD54L and/or siTOP3A, labeled with EdU, irradiated with 4 Gy, and then incubated with BrdU. Representative images of BrdU foci reflecting DNA repair synthesis in G2 cells are shown. (E) BrdU foci were enumerated in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. **P < 0.01 (two-tailed t test). (F) Model illustrating the regulation of HR sub-pathway usage by TOP3A in intrinsically ATRX-deficient U2OS cells. TOP3A acts as a positive SDSA-promoting HR factor together with RAD51AP1 and RAD54B to facilitate SDSA usage. Additionally, TOP3A indirectly inhibits the usage of the dHJ sub-pathway in ATRX-deficient cells. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns: not significant (two-tailed t test).

Journal: bioRxiv

Article Title: PARP inhibitor synthetic lethality reveals homologous recombination sub-pathway architecture

doi: 10.64898/2026.03.06.709881

Figure Lengend Snippet: (A) Representative images of γH2AX foci in U2OS ATRX G2 cells following transfection with the indicated siRNAs and subsequent irradiation are shown. (B) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs, and γH2AX foci were enumerated at 12 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. Striped bars indicate TOP3A depletion. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown was confirmed by western blotting. (C) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs. They were irradiated with 2 Gy, collected after 16h, and processed to obtain chromosome spreads. SCEs per spread were quantified and normalized to 70 chromosomes. Results from individual experiments, each derived from 40 cells, are indicated (left), and red horizontal lines indicate the mean. The bar graph shows the IR-induced SCE numbers (total SCEs after background subtraction) (right). IR-induced SCE data show mean ± SEM, and results from individual experiments are indicated. 120 spreads and >7,000 chromosomes per condition were analysed from three independent experiments. (D) U2OS cells were transfected with siCTRL, siRAD54L and/or siTOP3A, labeled with EdU, irradiated with 4 Gy, and then incubated with BrdU. Representative images of BrdU foci reflecting DNA repair synthesis in G2 cells are shown. (E) BrdU foci were enumerated in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. **P < 0.01 (two-tailed t test). (F) Model illustrating the regulation of HR sub-pathway usage by TOP3A in intrinsically ATRX-deficient U2OS cells. TOP3A acts as a positive SDSA-promoting HR factor together with RAD51AP1 and RAD54B to facilitate SDSA usage. Additionally, TOP3A indirectly inhibits the usage of the dHJ sub-pathway in ATRX-deficient cells. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns: not significant (two-tailed t test).

Techniques: Transfection, Irradiation, Expressing, Derivative Assay, Knockdown, Western Blot, Labeling, Incubation, Two Tailed Test

(A) HeLa and U2OS cells were transfected with the indicated siRNAs, irradiated with 2 Gy, and γH2AX foci were enumerated at 8 h (HeLa) or 10 h (U2OS) post-irradiation in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 2-3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown of HIRA was confirmed by western blotting (right). (B) HeLa pGC cells were transfected with the indicated siRNAs, followed by transfection with the I-SceI expression plasmid to induce site-specific DSBs. The percentage of GFP-positive cells, representing HR repair events, was measured by flow cytometry. All data show mean ± SEM (n = 3). Results from individual experiments are indicated. (C) U2OS cells were transfected with the siHIRA and RAD51 foci were enumerated 2 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 2). Results from individual experiments, each derived from 40 cells, are indicated. *P < 0.05; **P < 0.01; ***P < 0.001; ns: not significant (two-tailed t test).

Journal: bioRxiv

Article Title: PARP inhibitor synthetic lethality reveals homologous recombination sub-pathway architecture

doi: 10.64898/2026.03.06.709881

Figure Lengend Snippet: (A) HeLa and U2OS cells were transfected with the indicated siRNAs, irradiated with 2 Gy, and γH2AX foci were enumerated at 8 h (HeLa) or 10 h (U2OS) post-irradiation in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 2-3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown of HIRA was confirmed by western blotting (right). (B) HeLa pGC cells were transfected with the indicated siRNAs, followed by transfection with the I-SceI expression plasmid to induce site-specific DSBs. The percentage of GFP-positive cells, representing HR repair events, was measured by flow cytometry. All data show mean ± SEM (n = 3). Results from individual experiments are indicated. (C) U2OS cells were transfected with the siHIRA and RAD51 foci were enumerated 2 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 2). Results from individual experiments, each derived from 40 cells, are indicated. *P < 0.05; **P < 0.01; ***P < 0.001; ns: not significant (two-tailed t test).

Techniques: Transfection, Irradiation, Derivative Assay, Knockdown, Western Blot, Expressing, Plasmid Preparation, Flow Cytometry, Two Tailed Test

(A) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs, and γH2AX foci were enumerated at 12 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown was confirmed by western blotting. (B) U2OS DR-GFP reporter cells carry a GFP cassette that is disrupted by the I-SceI sequence and a second truncated GFP cassette. I-SceI-induced HR repair events result in the expression of functional GFP. Cells were transfected with the indicated siRNAs followed by the I-SceI plasmid transfection, and GFP-positive cells were measured by flow cytometry. All data show mean ± SEM (n = 3). Results from individual experiments are indicated. (C) Model illustrating the regulation of HR sub-pathway usage in intrinsically ATRX-deficient U2OS cells. In addition to RAD51AP1 and RAD54B, HIRA contributes to SDSA, but not to the dHJ sub-pathway, through its H3.3 deposition function. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns: not significant (two-tailed t test).

Journal: bioRxiv

Article Title: PARP inhibitor synthetic lethality reveals homologous recombination sub-pathway architecture

doi: 10.64898/2026.03.06.709881

Figure Lengend Snippet: (A) U2OS ATRX cells, with (blue) and without (green) doxycycline-induced ATRX expression, were transfected with the indicated siRNAs, and γH2AX foci were enumerated at 12 h following 2 Gy IR in EdU-negative G2 cells. Spontaneous foci were subtracted. All data show mean ± SEM (n = 3). Results from individual experiments, each derived from 40 cells, are indicated. Knockdown was confirmed by western blotting. (B) U2OS DR-GFP reporter cells carry a GFP cassette that is disrupted by the I-SceI sequence and a second truncated GFP cassette. I-SceI-induced HR repair events result in the expression of functional GFP. Cells were transfected with the indicated siRNAs followed by the I-SceI plasmid transfection, and GFP-positive cells were measured by flow cytometry. All data show mean ± SEM (n = 3). Results from individual experiments are indicated. (C) Model illustrating the regulation of HR sub-pathway usage in intrinsically ATRX-deficient U2OS cells. In addition to RAD51AP1 and RAD54B, HIRA contributes to SDSA, but not to the dHJ sub-pathway, through its H3.3 deposition function. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns: not significant (two-tailed t test).

Techniques: Expressing, Transfection, Derivative Assay, Knockdown, Western Blot, Sequencing, Functional Assay, Plasmid Preparation, Flow Cytometry, Two Tailed Test

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

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