ogg1 inhibitor th5487  (Tocris)

Journal: Frontiers in Cell and Developmental Biology

Article Title: OGG1 competitive inhibitors show important off-target effects by directly inhibiting efflux pumps and disturbing mitotic progression

doi: 10.3389/fcell.2023.1124960

Figure Lengend Snippet: The addition of OGG1 competitive inhibitors TH5487 and SU0268 to the medium results in higher staining with fluorescent dyes. (A) U2OS cells were stained with mitochondrial markers MitoTracker Green and TMRE in the presence of 10 µM of OGG1 inhibitors TH5487 or SU0268. Control cells were stained in DMEM containing DMSO at .1%. Fluorescence was acquired in a plate reader. Fluorescent intensity for each of the channels was normalized to the values measured in cells stained in control cells. Ploted values correspond to at least three technical replicates from four independent experiments. Statistical analysis was performed with a Kruskal-Wallis test. (****) p < 0.0001; (**) p < 0.005. (B) U2OS cells were stained with NucLight in the presence or absence of OGG1 inhibitors TH5487 and SU0268 combined or not with Verapamil (A) and imaged with the Incucyte ® Live-Cell Analysis System. (C) OGG1 KO cells generated by CRISPR/Cas9 were validated by Western Blot analysis using an antibody against OGG1. Vinculine was used as a loading control. Clones 2.1 and 3.3 were selected and used in further experiments. (D) U2OS WT and OGG1 KO cells, were stained with NucLight and Picogreen in the presence or absence of OGG1 inhibitors or Verapamil (all used at 10 µM) and imaged with a high-content imaging microscope in both the green and the red channels. Representative images used for the quantification are presented in Figure S1. The graph corresponds to the NucLight fluorescence intensity measured in the nucleus for up to 7000 cells for each condition. One representative experiment out of three is presented. Statistical analysis was performed with a Kruskal-Wallis test. (****) p < 0.0001. (E) U2OS WT and OGG1 KO cells were stained with SiR-DNA in the presence of different concentrations of TH5487 and SU0268 (1, 2.5, 5, 10, 25 or 50 µM) and analysed by flow-cytometry. Results from a representative experiment out of three are presented. The fluorescence distributions of cells stained in the presence of different concentration of TH5487 or SU0268 were compared to control cells on FlowJo using a chi-squared test (****) p -value < 0.001). (F) U2OS WT and OGG1 KO cells (clone 2.1) were stained with SiR-DNA or TMRE in the presence of different concentrations of TH5487 and SU0268 (1, 2.5, 5, 10, 25 or 50 µM) and analysed by flow-cytometry. The median of at least three independent experiments is shown. Statistical analysis was performed with Ordinary one-way ANOVA (*) p < 0.05; (**) p < 0.01; (***) p < 0.001; (****) p < 0.0001.

Article Snippet: OGG1 inhibitors TH5487 (M9506) and SU0268 (HY-139056 (MedChemExpress); 10-4700 (Focus Biomolecules) and MBS5765223 (MyBiosource)) were purchased from Clinisciences.

Techniques: Staining, Control, Fluorescence, Cell Analysis, Generated, CRISPR, Western Blot, Clone Assay, Imaging, Microscopy, Flow Cytometry, Concentration Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: OGG1 competitive inhibitors show important off-target effects by directly inhibiting efflux pumps and disturbing mitotic progression

doi: 10.3389/fcell.2023.1124960

Figure Lengend Snippet: OGG1 inhibitors TH5487 and SU0268 inhibit the efflux pump activity of BCRP, MDR1 and MRP1. (A) Scheme illustrating the vesicle transport assay. See results and material and methods sections for details. (B) Vesicles expressing inside-out BCRP, MDR1 or MRP1 were incubated with the corresponding selective inhibitors (Kol143 0.2 µM—Valspodar 1 µM–Benzbromarone 200 µM), TH5487 and SU0268 (10 and 100 µM) and the specific pump substrates (E3S-NMQ-E217βG for BCRP, MDR1 or MRP1 respectively). In the graph, the substrate levels inside the vesicles are expressed as percentage of the ones in DMSO control and referred as relative pump inhibition. The means and the s.d. of three replicates are shown. The statistical analysis was performed with One-way ANOVA (*) p < 0.05; (**) p < 0.01; (***) p < 0.001; (****) p < 0.0001. (C) The expression of levels of BCRP and MDR1 were quantified by RT-qPCR in the indicated cell lines (HMLE, A549, and SH-SY5Y). The table reports the normalized transcript expression values as indicated in the Protein Atlas (nTPM). (D) SH-SY5Y and HMLE cells were stained with SiR-DNA in the presence of different concentrations of TH5487, SU0268 and the efflux pump inhibitors Verapamil (Vera) and Zosuquidar/LY-335979 (LY). (E) A549 and HMLE cells were stained with HO33342 in the presence of different concentrations of TH5487, SU0268 and the efflux pump inhibitors Verapamil (Vera) and Fumitremorgin C (Fumi) (F) A549 cells were exposed to 1 µM of Mitoxantrone (MTX) in the presence of different concentrations of TH5487, SU0268 and the efflux pump inhibitors Verapamil (Vera) and Fumitremorgin (Fumi). For (D–F) , the median fluorescence intensity was normalized to the median of the cells stained without inhibitors (NT) and data presented as a mean±sd for three independent experiments. Statistical analysis was performed with Ordinary one-way ANOVA (*) p < 0.05; (**) p < 0.01; (***) p < 0.001; (****) p < 0.0001.

Article Snippet: OGG1 inhibitors TH5487 (M9506) and SU0268 (HY-139056 (MedChemExpress); 10-4700 (Focus Biomolecules) and MBS5765223 (MyBiosource)) were purchased from Clinisciences.

Techniques: Activity Assay, Vesicular Transport Assay, Expressing, Incubation, Control, Inhibition, Quantitative RT-PCR, Staining, Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: OGG1 competitive inhibitors show important off-target effects by directly inhibiting efflux pumps and disturbing mitotic progression

doi: 10.3389/fcell.2023.1124960

Figure Lengend Snippet: OGG1 inhibitors TH5487 and SU0268 increase the effects of the chemotherapeutical drug Etoposide, independently of OGG1. (A) U2OS WT and OGG1 KO cells were exposed to Etoposide alone or in combination with 10 µM of TH5487, SU0268 or Verapamil. Immediately after exposure to the drugs, cells were fixed and DSB were visualized by immunofluorescence using an antibody against γH2AX (green). Nuclei were stained with Hoechst 33342 (blue). Statistical analysis was performed with a Kruskal Wallis test. A segmentation algorithm was used to quantify γH2AX foci number in at least 1000 cells for each condition. Scale bar 50 µm. (B) Distribution of γH2AX foci number per nuclei is presented for a representative experiment out of three. Statistic analysis was performed with a Kruskal Wallis test (***) p < 0.001; (****) p < 0.0001. (C) The number of γH2AX foci per nuclei was normalized to the number of foci in cells exposed to Etoposide and the mean from three independent experiments is represented in the graph. Statistical analysis was performed with a Kruskal Wallis test (**) p < 0.01; (***) p < 0.001; (****) p < 0.0001. (D) Cell survival was evaluated in U2OS WT and OGG1 KO cells, 4 days after exposure to a single treatment with Etoposide or a combined treatment with Etoposide and 10 µM of TH5487, SU0268 or Verapamil. The number of cells were normalized to the untreated control cells. The mean of three independent experiments is represented in the graph. Statistical analysis was performed with a Kruskal Wallis test (*) p < 0.05; (**) p < 0.01; (***) p < 0.001; (****) p < 0.0001.

Article Snippet: OGG1 inhibitors TH5487 (M9506) and SU0268 (HY-139056 (MedChemExpress); 10-4700 (Focus Biomolecules) and MBS5765223 (MyBiosource)) were purchased from Clinisciences.

Techniques: Immunofluorescence, Staining, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: OGG1 competitive inhibitors show important off-target effects by directly inhibiting efflux pumps and disturbing mitotic progression

doi: 10.3389/fcell.2023.1124960

Figure Lengend Snippet: OGG1 inhibitor SU0268 impairs mitotic progression in an OGG1 independent way. (A) The proliferation of U2OS WT and OGG1 KO cells under chronic treatment with TH5487 and SU0268 (10 µM) was followed with Incucyte for one 1 week as described in materials and methods. Data are presented as mean confluence ±s.d. of at least three technical replicates from three independent experiments (left side). Mixed-effect analysis was performed (****) p < 0.0001. (B) Representative Incucyte transmission images of U2OS WT and OGG1 KO2 cells at basal conditions or treated with SU026. Scale bar 50 µm (C) Cells were exposed to Nocodazole or SU0268 for 6 h, DNA was stained with Ho33342 and cells were imaged by fluorescence microscopy. For the untreated cells, the different phases of mitosis (Prophase, Metaphase, Anaphase and Telophase) are illustrated. Only cells in Prophase were observed upon exposure to SU0268 or Nocodazole. Scale bars 10 µm. (D) Cells undergoing mitosis were quantified by flow cytometry using an antibody against Phospho-H3. A higher accumulation of cells in mitosis was observed for both U2OS WT and OGG1 KO2 in cells exposed to SU0268 compared to untreated cells or cells exposed to TH5487.

Article Snippet: OGG1 inhibitors TH5487 (M9506) and SU0268 (HY-139056 (MedChemExpress); 10-4700 (Focus Biomolecules) and MBS5765223 (MyBiosource)) were purchased from Clinisciences.

Techniques: Transmission Assay, Staining, Fluorescence, Microscopy, Flow Cytometry

Journal: Frontiers in Cell and Developmental Biology

Article Title: OGG1 competitive inhibitors show important off-target effects by directly inhibiting efflux pumps and disturbing mitotic progression

doi: 10.3389/fcell.2023.1124960

Figure Lengend Snippet: Cellular effects of the OGG1 inhibitors TH5487 and SU0268. Beside their activity as inhibitors of the DNA glycosylase activity of OGG1, TH5487 and SU0268 have several off-target effects that are independent on OGG1. Both molecules inhibit the activity of the efflux pumps MDR1 and BCRP1, thus increasing the intracellular concentration of fluorescent dyes and chemotherapeutical molecules. In addition, SU0268 impairs mitotic progression.

Article Snippet: OGG1 inhibitors TH5487 (M9506) and SU0268 (HY-139056 (MedChemExpress); 10-4700 (Focus Biomolecules) and MBS5765223 (MyBiosource)) were purchased from Clinisciences.

Techniques: Activity Assay, Concentration Assay

Journal: Cells

Article Title: RELA∙8-Oxoguanine DNA Glycosylase1 Is an Epigenetic Regulatory Complex Coordinating the Hexosamine Biosynthetic Pathway in RSV Infection

doi: 10.3390/cells11142210

Figure Lengend Snippet: Quantitative RT-PCR primers.

Article Snippet: The selective OGG1 inhibitor TH5487 (SelleckChem, Houston, TX, USA) was used at 10 μM and added to the culture medium every 6–12 h. The generation of hSAEC expressing doxycycline (Dox)-inducible RelA shRNA was described earlier [ ].

Techniques: Quantitative RT-PCR

Journal: Cells

Article Title: RELA∙8-Oxoguanine DNA Glycosylase1 Is an Epigenetic Regulatory Complex Coordinating the Hexosamine Biosynthetic Pathway in RSV Infection

doi: 10.3390/cells11142210

Figure Lengend Snippet: OGG1 regulates the expression of core genes in the HBP. ( A ) Scrambled (Scr) or OGG1-targeting siRNA was transfected into hSAECs. Cells were mock- or RSV-infected (MOI = 1, for 24 h), and Q-RT-PCR was performed for GFPT2 expression. Data are presented as fold change untreated controls normalized to PPIA. ( B ) hSAECs were mock- or RSV-infected in the absence (DMSO) or presence of TH5487. Shown are the fold change of GFPT2 mRNA. ( C ) A549 epithelial carcinoma cells were treated as in ( B ). ( D ) Wild-type or OGG1 −/− cells (by CrispR/Cas9) were mock-or RSV-infected. Shown is the fold change in GFPT2 mRNA expression. ( E ) GNPNAT1 mRNA expression in RSV-infected cells treated with solvent (DMSO) or TH5487. ( F ) UAP1 mRNA levels in cells treated as in panel ( E ). *, p <0.05; **, p < 0.01 post-hoc Tukey’s test.

Article Snippet: The selective OGG1 inhibitor TH5487 (SelleckChem, Houston, TX, USA) was used at 10 μM and added to the culture medium every 6–12 h. The generation of hSAEC expressing doxycycline (Dox)-inducible RelA shRNA was described earlier [ ].

Techniques: Expressing, Transfection, Infection, Reverse Transcription Polymerase Chain Reaction, CRISPR, Solvent

Journal: Cells

Article Title: RELA∙8-Oxoguanine DNA Glycosylase1 Is an Epigenetic Regulatory Complex Coordinating the Hexosamine Biosynthetic Pathway in RSV Infection

doi: 10.3390/cells11142210

Figure Lengend Snippet: OGG1 is required for RSV-induced GFPT2 expression. ( A ) hSAECs were treated with solvent (DMSO) or TH5487 and mock- or RSV-infected as indicated. Cells were stained with anti-GFPT2 Ab (red) and nuclei by DAPI (blue). Bar indicates 30 μm. ( B ) The quantitation of GFPT2 staining. GFPT2 fluorescence was segmented by StarDist and plotted as arbitrary fluorescence/cell for each treatment condition. **, p < 0.01 by post-hoc Tukey comparison.

Article Snippet: The selective OGG1 inhibitor TH5487 (SelleckChem, Houston, TX, USA) was used at 10 μM and added to the culture medium every 6–12 h. The generation of hSAEC expressing doxycycline (Dox)-inducible RelA shRNA was described earlier [ ].

Techniques: Expressing, Solvent, Infection, Staining, Quantitation Assay, Fluorescence, Comparison

Journal: Cells

Article Title: RELA∙8-Oxoguanine DNA Glycosylase1 Is an Epigenetic Regulatory Complex Coordinating the Hexosamine Biosynthetic Pathway in RSV Infection

doi: 10.3390/cells11142210

Figure Lengend Snippet: RSV induces phospho-Ser536 RELA binding to OGG1. ( A ) Co-immunoprecipitation (Co-IP) experiments. hSAECs lysates from mock- or RSV-infected cells were IPed with anti-OGG1 antibody. The immune complexes were fractionated and subjected to IB with anti-RELA. Arrowhead shows the 65 kDa RELA. ( B ) Co-IP of mock- or RSV-infected cells with anti-RELA Ab. Immune complexes were fractionated and subjected to IB with anti-OGG1. Arrowhead indicates the 50 kDa OGG1. ( C ) Lysates were prepared from FLAG-M2-OGG1-expressing A549 cells after various times of RSV infection and IPed with anti-FLAG M2 beads. Top panel, immune complexes were stained with anti-phospho-Ser536 RELA. The blot was stripped and re-probed to confirm equivalent levels of FLAG-OGG1 capture. Bottom panels are input showing total RELA and b actin. Molecular-weight markers are kDa (kD).

Article Snippet: The selective OGG1 inhibitor TH5487 (SelleckChem, Houston, TX, USA) was used at 10 μM and added to the culture medium every 6–12 h. The generation of hSAEC expressing doxycycline (Dox)-inducible RelA shRNA was described earlier [ ].

Techniques: Binding Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Infection, Expressing, Staining, Molecular Weight

Journal: Cells

Article Title: RELA∙8-Oxoguanine DNA Glycosylase1 Is an Epigenetic Regulatory Complex Coordinating the Hexosamine Biosynthetic Pathway in RSV Infection

doi: 10.3390/cells11142210

Figure Lengend Snippet: OGG1 inducibly binds to HBP regulatory elements and recruits phospho-Pol II. XChIP was performed on control or RSV-infected hSAECs; data are expressed as a fold change over IgG control. ( A ) OGG1 binding to the GFPT2 intragenic enhancer. ( B ) OGG1 binding to the UAP1 promoter. ( C – F ), Cells were RSV infected in the presence of solvent (DMSO) or TH5487 as indicated. ( C , D ), XChIP for RELA binding. ( E , F ), XChIP for phospho-Ser2 CTD Pol II binding. *, p < 0.05; **, p < 0.01; n.s., not significant. Each symbol is an independent immunoprecipitation.

Article Snippet: The selective OGG1 inhibitor TH5487 (SelleckChem, Houston, TX, USA) was used at 10 μM and added to the culture medium every 6–12 h. The generation of hSAEC expressing doxycycline (Dox)-inducible RelA shRNA was described earlier [ ].

Techniques: Control, Infection, Binding Assay, Solvent, Immunoprecipitation

Journal: Cells

Article Title: RELA∙8-Oxoguanine DNA Glycosylase1 Is an Epigenetic Regulatory Complex Coordinating the Hexosamine Biosynthetic Pathway in RSV Infection

doi: 10.3390/cells11142210

Figure Lengend Snippet: Graphical summary. Schematic of effects of RSV-induced oxidative stress on the formation of a RELAOGG1 complex that binds to nucleosome-free domains in the HBP regulatory regions. OGG1 activity is required for the RELA recruitment and association of the transcriptional elongation-competent form of phosphorylated RNA Pol II. HBP enzymes GFPT1/2, GNPNAT1, PGM3 and UAP1 catalyze the conversion of fructose-6 phosphate into UDP-GlcNAc, required for N-linked glycosylation of integrins and basal lamina proteins.

Article Snippet: The selective OGG1 inhibitor TH5487 (SelleckChem, Houston, TX, USA) was used at 10 μM and added to the culture medium every 6–12 h. The generation of hSAEC expressing doxycycline (Dox)-inducible RelA shRNA was described earlier [ ].

Techniques: Activity Assay, Glycoproteomics

Journal: The FASEB Journal

Article Title: Enzymatically inactive OGG1 binds to DNA and steers base excision repair toward gene transcription

doi: 10.1096/fj.201902243R

Figure Lengend Snippet: Binding and enzymatic activity of OGG1 mutant's. A and B, Binding of wt and mutant OGG1's to 8‐oxoG‐containing probes. Purified recombinant proteins (20, 10, and 5 nM) were incubated with 100 fmol 8‐oxoG‐containing Cy5‐labeled DNA oligos (total volume is 10 µL) for 10 minutes on ice. Retardation of protein/DNA complex was analyzed by EMSA. The right panels show quantification of the corresponding experiments. C, Changes in amino acids crucial for enzymatic function decreases/prevents base excision activity of OGG1. Purified recombinant proteins as described above were mixed with 100 fmol 8‐oxoG‐containing Cy5‐labeled DNA oligos for 10 minutes at room temperature. The cleaved fragments were separated from the intact strand in a 20% of polyacrylamide gel. DNA bands were visualized by using a LI‐COR Odyssey CLx system. The right panels show quantification of the corresponding experiments. D, Equal concentrations and titrations of OGG1 proteins were used in the above Figures. All experiments were performed three times. Quantification analyses were performed using Image J and the data are presented as the means ± the standard error. A two‐way analysis of variance (ANOVA) was applied to determine the significance of the difference. *** P < .001, compared with GST in (A), and with wt OGG1 in (B) and (C)

Article Snippet: OGG1 inhibitor 4‐(4‐Bromo‐2‐oxo‐3H‐benzimidazol‐1‐yl)‐N‐(4‐iodophenyl) piperidine‐1‐carboxamide (TH5487, Cat # PC‐35806) was purchased from ProbeChem (Shanghai, China).

Techniques: Binding Assay, Activity Assay, Mutagenesis, Purification, Recombinant, Incubation, Labeling

Journal: The FASEB Journal

Article Title: Enzymatically inactive OGG1 binds to DNA and steers base excision repair toward gene transcription

doi: 10.1096/fj.201902243R

Figure Lengend Snippet: Binding to the DNA substrate but not base excision activity is required for OGG1 to modulate pro‐inflammatory gene expression. Ogg1 −/− MEF cells were transfected with plasmids expressing Flag‐tagged (A) or YFP‐tagged (B) wt OGG1 or site‐specific mutants, and then, exposed to 20 ng/mL TNFα for 0, 30, 60, and 120 minutes. Cxcl 2 gene expression was determined by real‐time PCR. The lower panels of A and B show the transfection efficiencies of OGG1 and mutants. Changes in gene expression in cells transfected with control plasmids PcDNA 3.1 (+) (A) and YFP (B) were taken as control. All experiments were performed three times. The data are presented as the means ± the standard error. A two‐way analysis of variance (ANOVA) was applied to determine the significance of the difference. *** P < .001, ns: no significance, compared with PcDNA3.1 in (A) and with YFP in (B)

Article Snippet: OGG1 inhibitor 4‐(4‐Bromo‐2‐oxo‐3H‐benzimidazol‐1‐yl)‐N‐(4‐iodophenyl) piperidine‐1‐carboxamide (TH5487, Cat # PC‐35806) was purchased from ProbeChem (Shanghai, China).

Techniques: Binding Assay, Activity Assay, Gene Expression, Transfection, Expressing, Real-time Polymerase Chain Reaction, Control

Journal: The FASEB Journal

Article Title: Enzymatically inactive OGG1 binds to DNA and steers base excision repair toward gene transcription

doi: 10.1096/fj.201902243R

Figure Lengend Snippet: Expression of Cxcl2 in wt and OGG1 variant‐expressing cells as shown by FISH Ogg1 −/− MEF cells were transfected with plasmids expressing A, YFP, B, wt OGG1, C, OGG1 K249Q, D) OGG1 C253A, and then, exposed to 20 ng/mL TNFα for 0, 30, 60, and 120 minutes. The levels of Cxcl 2 mRNA were determined by fluorescence in situ hybridization (red). The nuclei of cells were counter stained with DAPI (red: Cxcl 2 mRNA; blue: DNA; green: OGG1). Bar, 10 μm. A representative result of three experiments is shown

Article Snippet: OGG1 inhibitor 4‐(4‐Bromo‐2‐oxo‐3H‐benzimidazol‐1‐yl)‐N‐(4‐iodophenyl) piperidine‐1‐carboxamide (TH5487, Cat # PC‐35806) was purchased from ProbeChem (Shanghai, China).

Techniques: Expressing, Variant Assay, Transfection, Fluorescence, In Situ Hybridization, Staining

Journal: The FASEB Journal

Article Title: Enzymatically inactive OGG1 binds to DNA and steers base excision repair toward gene transcription

doi: 10.1096/fj.201902243R

Figure Lengend Snippet: Recruitment of enzymatically inactive OGG1 to the promoter facilitates transcriptional activation of pro‐inflammatory genes. A, Enrichment of wt OGG1 and the mutants on the CXCL1 promoter in response to TNFα exposure. HEK 293 cells were transfected with Flag‐tagged wt OGG1 or OGG1 mutants, and then, mock treated or TNF‐exposed for 60 minutes. Enrichment of wt OGG1 and mutants on the CXCL1 promoter was determined by ChIP‐coupled qPCR (left panel). The right panel shows equal expression of each OGG1 in transfected cells. B, TNFα exposure induces oxidative modification of OGG1 at cysteine residues. YFP or YFP‐wt OGG1‐expressing cells were mock‐treated or NAC (10 mM)‐treated for 60 minutes, and then, exposed to TNFα for 60 minutes. Whole cell extracts were made in the presence of DCP‐Bio1 (a cysteine sulfenic acid‐reacting agent). Immunoprecipitation with YFP beads was conducted and levels of DCP‐Bio1‐tagged YFP‐wt OGG1 were determined by western blot. C, The effects of OS and inhibitors on OGG1 DNA interactions. HEK 293 cells were transfected with Flag‐tagged wt OGG1, and then, exposed to TNFα for 60 minutes with or without pretreatment of NAC (10 mM), TH5487 (10 µM), or O8 (10 µM). ChIP was performed using antibody to Flag‐tag. PCR amplification of the TSS adjacent CXCL1 promoter region was performed to determine the enrichment of OGG1. D, The effects of oxidative inactivation or inhibitors on CXCL1 gene expression. HEK 293 cells were exposed to TNFα for 60 minutes with or without pretreatment of NAC, TH5487, or O8. CXCL1 gene expression was measured by real‐time qPCR. E, The roles of wt OGG1 and the mutants in transcription activation were analyzed using a dual reporter assay. Ogg1 −/− MEF cells expressing wt OGG1 or the mutants were co‐transfected with a luciferase dual reporter system. Firefly luciferase mRNA expression is driven by the Cxcl 2 promoter. Cells were mock‐treated or exposed to TNFα for 0, 30, 60, and 120 minutes. The mRNA levels of firefly luciferase were assessed by real‐time qPCR. All experiments were performed three times. The data are presented as the means ± the standard error. A two‐way analysis of variance indicated the significance of the difference. * P < .05 and *** P < .001, ns: no significance, compared with PcDNA3.1

Article Snippet: OGG1 inhibitor 4‐(4‐Bromo‐2‐oxo‐3H‐benzimidazol‐1‐yl)‐N‐(4‐iodophenyl) piperidine‐1‐carboxamide (TH5487, Cat # PC‐35806) was purchased from ProbeChem (Shanghai, China).

Techniques: Activation Assay, Transfection, Expressing, Modification, Immunoprecipitation, Western Blot, FLAG-tag, Amplification, Gene Expression, Reporter Assay, Luciferase

Journal: The FASEB Journal

Article Title: Enzymatically inactive OGG1 binds to DNA and steers base excision repair toward gene transcription

doi: 10.1096/fj.201902243R

Figure Lengend Snippet: Substrate binding is required for OGG1‐dependent pro‐inflammatory gene expression. ROS generate 8‐oxoG in the promoter region. A, Oxidative inactivation, posttranslational modifications or mutations that halt enzymatic activity but allow binding and pre‐excision steps by OGG1 are exploited to promote gene expression. B, Mutation (s) or inhibitors of OGG1 that blocks substrate engagement inhibits OGG1 function in gene expression

Article Snippet: OGG1 inhibitor 4‐(4‐Bromo‐2‐oxo‐3H‐benzimidazol‐1‐yl)‐N‐(4‐iodophenyl) piperidine‐1‐carboxamide (TH5487, Cat # PC‐35806) was purchased from ProbeChem (Shanghai, China).

Techniques: Binding Assay, Gene Expression, Activity Assay, Mutagenesis