Journal: Molecular Medicine

Article Title: Inhibition of OGG1 ameliorates pulmonary fibrosis via preventing M2 macrophage polarization and activating PINK1-mediated mitophagy

doi: 10.1186/s10020-024-00843-6

Figure Lengend Snippet: TH5487 mitigates BLM-induced pulmonary fibrosis and M2 macrophage polarization, which is partly hindered by Mdivi-1. A BLM-administrated mice were treated with TH5487 or co-treated with TH5487 and Mdivi-1. HE staining and Masson staining were performed to observe the histological changes. B The expression level of Collagen I and α-SMA in lung tissues was examined by western blot. C - D Immunofluorescent analysis was used to detect CD206-positive (M2) cells. ***p < 0.001 vs Sham; ###p < 0.001 vs BLM; &p < 0.05, &&&p < 0.001 vs BLM + TH5487

Article Snippet: One day after BLM induction, the pulmonary fibrosis mice were treated intraperitoneally with TH5487 (30 mg/kg/twice one week; OGG1-specific inhibitor, Selleck) in BLM + TH5487 group, and were treated intraperitoneally with TH5487 and Mitochondrial division inhibitor 1 (Mdivi-1; 10 mg/kg/twice one week; Selleck) in BLM + TH5487 + Mdivi-1 group.

Techniques: Staining, Expressing, Western Blot

Journal: Molecular Medicine

Article Title: Inhibition of OGG1 ameliorates pulmonary fibrosis via preventing M2 macrophage polarization and activating PINK1-mediated mitophagy

doi: 10.1186/s10020-024-00843-6

Figure Lengend Snippet: TH5487 mitigates BLM-induced oxidative stress and mitophagy of mice, which is partly hindered by Mdivi-1. A - C BLM-administrated mice were treated with TH5487 or co-treated with TH5487 and Mdivi-1. The levels of 4-NHE, MDA and ROS of lung tissues was examined using their corresponding commercial kits. D Representative images of mtROS production in lung OCT sections. E Representative images of LC3B-MitoTracker double immunofluorescent staining of lung tissues. F The expression level of PINK1 and Parkin in lung tissues was examined by western blot. ***p < 0.001 vs Sham; ###p < 0.001 vs BLM; &p < 0.05, &&p < 0.01, &&&p < 0.001 vs BLM + TH5487

Article Snippet: One day after BLM induction, the pulmonary fibrosis mice were treated intraperitoneally with TH5487 (30 mg/kg/twice one week; OGG1-specific inhibitor, Selleck) in BLM + TH5487 group, and were treated intraperitoneally with TH5487 and Mitochondrial division inhibitor 1 (Mdivi-1; 10 mg/kg/twice one week; Selleck) in BLM + TH5487 + Mdivi-1 group.

Techniques: Staining, Expressing, Western Blot

Journal: Molecular medicine (Cambridge, Mass.)

Article Title: Inhibition of OGG1 ameliorates pulmonary fibrosis via preventing M2 macrophage polarization and activating PINK1-mediated mitophagy.

doi: 10.1186/s10020-024-00843-6

Figure Lengend Snippet: Fig. 2 TH5487 mitigates BLM-induced pulmonary fibrosis and M2 macrophage polarization, which is partly hindered by Mdivi-1. A BLM-administrated mice were treated with TH5487 or co-treated with TH5487 and Mdivi-1. HE staining and Masson staining were performed to observe the histological changes. B The expression level of Collagen I and α-SMA in lung tissues was examined by western blot. C-D Immunofluorescent analysis was used to detect CD206-positive (M2) cells. ***p < 0.001 vs Sham; ###p < 0.001 vs BLM; &p < 0.05, &&&p < 0.001 vs BLM + TH5487

Article Snippet: One day after BLM induction, the pulmonary fibrosis mice were treated intraperitoneally with TH5487 (30 mg/ kg/twice one week; OGG1-specific inhibitor, Selleck) in BLM + TH5487 group, and were treated intraperitoneally with TH5487 and Mitochondrial division inhibitor 1 (Mdivi-1; 10 mg/kg/twice one week; Selleck) in BLM + TH5487 + Mdivi-1 group.

Techniques: Staining, Expressing, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: 8-Oxoguanine DNA glycosylase 1 selectively modulates ROS-responsive NF-κB targets through recruitment of MSK1 and phosphorylation of RelA/p65 at Ser276

doi: 10.1016/j.jbc.2023.105308

Figure Lengend Snippet: ROS selectively regulates NF-κB–driven inflammatory gene expression . A – J , HEK293 cells were exposed to TNFα (20 ng/ml) for 1 h with or without pretreatment of BMS-345541 (10 μM) or NAC (10 mM). CXCL1 , CXCL2 , CXCL8 , CCL2 , CCL5 , CCL20 , TNF , NFKBIA , NFKBIB , and NFKBIE gene expression was measured by real-time qPCR. All experiments were performed five times. Data are presented as mean ± SD. ∗∗ p < 0.01, ns, not significant. NAC, N-acetyl-L-cysteine; qPCR, quantitative PCR; ROS, reactive oxygen species.

Article Snippet: Cells were treated with or without IKK inhibitor BMS-345541 (Sigma, Cat# B9935), 10 mM of NAC (Sigma, Cat# A7250), 10 μM of OGG1 inhibitor TH5487 (ProbeChem, Cat#, PC-35806),7,8-Dihydro-8-oxoguaninetriphosphatase inhibitor TH588 (Sigma, Cat# 5309160001), TH2840 (Provided by Dr T. Helleday, Karolinska Institute, Stockholm, Sweden;) for 1 h, followed by exposure to 20 ng/ml TNFα (Sigma, Cat# H8961) for various durations of time.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Journal of Innate Immunity

Article Title: Innate Immune Responses to RSV Infection Facilitated by OGG1, an Enzyme Repairing Oxidatively Modified DNA Base Lesions

doi: 10.1159/000524186

Figure Lengend Snippet: OGG1 knockdown, siRNA depletion or inhibition of OGG1 substrate binding decreased RSV-induced proinflammatory gene expressions. a Schematic depiction of OGG1 knockdown by CRISP/Cas9 technology. Lack of OGG1 expression as shown by Western blotting ( b ) and microscopic imaging ( c ). Nuclei of cells were counter stained with DAPI. Images were photographed using an OLYMPUS Microscope System as in Fig. . Magnification, 20x lens and 3.6x (camera). Scale bar, 20 μm. d OGG1 KO (OGG1 − ) hSAECs expressed significantly lower levels of TNF, CCL20, IL6, CCL5, and CXCL10 in response to RSV infection. e SiRNA depletion of OGG1 but not NEIL1 or MTH1 decreased proinflammatory gene expression ( TNF, CCL20, IL6, CCL5, and CXCL10 ) in hSAECs. Lower panels, extent of OGG1, NEIL1, and MTH1 silencing as determined by Western blotting and qRT-PCR. siNT, non-targeting siRNA. f Inhibition of OGG1 substrate binding by TH5487 decreased inflammatory gene expression. The O8, inactive analog of TH5487, TH2480 had no effect. In ( b–d ), Cells were RSV-infected (MOI = 1) and total RNAs were isolated at 24 h. Data are mean ± SEM. *** p < 0.001; ** p < 0.01. NEIL1, human ortholog of E. coli Nei; MTH1, hMTH1; TH5487, 4-(4 bromo-2-oxo 3H-benzimidazol-1-yl)-N-(4-iodophenyl)piperidine-1-carboxamide; O8 (3,4-dichloro-benzo[b]thiophene-2-carboxylic acid hydrazide). In ( b − d ); TNF, CCL20, IL6, CCL5, and CXCL10 as in legend to Fig. . hMTH1, human homolog of E. Coli MutT 1. DAPI, 4,6-diamidino-2-phenylindole.

Article Snippet: A549 (ATCC, CCL-185) and MLE-12 (ATCC CRL-2110) cells were grown at 37°C and 5% CO 2 in DMEM-F-12 (1:1, Gibco) containing 10% fetal bovine serum (Gibco, Life Technologies, Inc), 100 units/mL penicillin (Gibco), and 100 µg/mL streptomycin (Gibco). hSAEC and CRISPR/Cas9-edited OGG1 knockout (KO) cells were cultured in small airway epithelial cell basal medium (Promo Cell C-21270), supplemented with supplement pack (C-39170) containing 0.004 mL/mL bovine pituitary, 10 ng/mL extract epidermal growth factor (recombinant human), 5 μg/mL insulin (recombinant human), 0.5 μg/mL hydrocortisone epinephrine, 0.5 μg/mL tri-iodo-L-thyronine, 6.7 ng/mL transferrin (recombinant human), 10 μg/mL retinoic acid, 0.1 ng/mL bovine serum albumin-fatty acid free (BSA-FAF) 2.5 mg/mL; AP endonuclease1' endonuclease activity inhibitor (Cat# CRT0044876; Sigma-Aldrich); TH5487 (4-(4-bromo-2-oxo-3H-benzimidazol-1-yl)-N-(4-iodophenyl)piperidine-1-carboxamide (Cat # HY-125276, Sigma Aldrich; TH2480 (inactive analog of TH5487) is provided by Dr. T. Helleday, Karolinska Institute, Stockholm, Sweden; O8 (Cat# SML1697; Sigma); deferoxamine (Cat# D9533, Sigma); anti-phospho NFκB/RelA/p65 (phosphoserine276) antibody (Ab) (Cat # ab106129); β-actin (Cat# 4970S, CST); anti-FLAG (Cat #F1804, Sigma-Aldrich); NFκB-RelA/p65 (D14E12) (Cat# 8284S, CST), OGG1 (Item #: ENZ-253, ProSpec); Ab to OGG1 (Novus, Cat# NB100-106), Ab to 8-oxo(d)Gua (Millipore Sigma, Cat# MAB3560).

Techniques: Inhibition, Binding Assay, Expressing, Western Blot, Imaging, Staining, Microscopy, Infection, Quantitative RT-PCR, Isolation