Journal: Iranian Journal of Parasitology

Article Title: Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice

doi: 10.18502/ijpa.v19i4.17163

Figure Lengend Snippet: Modulation of OGG1 activity during malaria infection

Article Snippet: The TH5487 (catalogue no. 6749) and O8-OGG1 (catalogue no. 6236) OGG1 inhibitors were purchased from Tocris Bioscience, USA.

Techniques: Activity Assay, Control, Sterility, Injection

Journal: Iranian Journal of Parasitology

Article Title: Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice

doi: 10.18502/ijpa.v19i4.17163

Figure Lengend Snippet: Comparative mean percentage total RBC count curves for the control mice and P. berghei - infected mice that received vehicle (VEH), TH5487 and O8 treatments. Data were analyzed using Unpaired Student’s t -test at P< 0.05 between malaria untreated mice and each of the treatment groups. Results were presented as Mean ± SEM (n= 8). The (*) denotes a comparison between the vehicle (VEH) and O8-treated mice and (¥) denotes a comparison between the vehicle (VEH) and TH5487-treated mice. C + VEH= Control + Vehicle; M + VEH= Malaria + Vehicle; M +TH5487= Malaria + TH5487; M + O8= Malaria + O8

Article Snippet: The TH5487 (catalogue no. 6749) and O8-OGG1 (catalogue no. 6236) OGG1 inhibitors were purchased from Tocris Bioscience, USA.

Techniques: Control, Infection, Comparison

Journal: Iranian Journal of Parasitology

Article Title: Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice

doi: 10.18502/ijpa.v19i4.17163

Figure Lengend Snippet: Effect of modulating OGG1 activity on serum TNF-α concentration on day 5 post P. berghei inoculation. C + VEH= Control + Vehicle; M + VEH= Malaria + Vehicle; M +TH5487= Malaria + TH5487; M + O8= Malaria + O8. Data are mean ± SEM, (n=6), * P< 0.05, *** P< 0.001

Article Snippet: The TH5487 (catalogue no. 6749) and O8-OGG1 (catalogue no. 6236) OGG1 inhibitors were purchased from Tocris Bioscience, USA.

Techniques: Activity Assay, Concentration Assay, Control

Journal: Iranian Journal of Parasitology

Article Title: Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice

doi: 10.18502/ijpa.v19i4.17163

Figure Lengend Snippet: Effect of modulating OGG1 activity on serum IFN-γ concentration on day 5 post P. berghei inoculation. C + VEH= Control + Vehicle; M + VEH= Malaria + Vehicle; M +TH5487= Malaria + TH5487; M + O8= Malaria + O8. Data are mean ± SEM, (n=6), * P< 0.05, ** P< 0.01

Article Snippet: The TH5487 (catalogue no. 6749) and O8-OGG1 (catalogue no. 6236) OGG1 inhibitors were purchased from Tocris Bioscience, USA.

Techniques: Activity Assay, Concentration Assay, Control

Journal: Iranian Journal of Parasitology

Article Title: Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice

doi: 10.18502/ijpa.v19i4.17163

Figure Lengend Snippet: Effect of modulating OGG1 activity on serum IL-2 concentration on day 5 post P. berghei inoculation. C + VEH= Control + Vehicle; M + VEH= Malaria + Vehicle; M +TH5487= Malaria + TH5487; M + O8= Malaria + O8. Data are mean ± SEM, (n=6), * P< 0.05

Article Snippet: The TH5487 (catalogue no. 6749) and O8-OGG1 (catalogue no. 6236) OGG1 inhibitors were purchased from Tocris Bioscience, USA.

Techniques: Activity Assay, Concentration Assay, Control

Journal: Iranian Journal of Parasitology

Article Title: Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice

doi: 10.18502/ijpa.v19i4.17163

Figure Lengend Snippet: Effect of modulating OGG1 activity on serum IL-10 concentration on day 5 post P. berghei inoculation. C + VEH= Control + Vehicle; M + VEH= Malaria + Vehicle; M +TH5487= Malaria + TH5487; M + O8= Malaria + O8. Data are mean ± SEM, (n=6), ** P< 0.01, *** P< 0.001

Article Snippet: The TH5487 (catalogue no. 6749) and O8-OGG1 (catalogue no. 6236) OGG1 inhibitors were purchased from Tocris Bioscience, USA.

Techniques: Activity Assay, Concentration Assay, Control

Journal: Iranian Journal of Parasitology

Article Title: Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice

doi: 10.18502/ijpa.v19i4.17163

Figure Lengend Snippet: Effect of modulating OGG1 activity on serum IL-6 concentration on day 5 post P. berghei inoculation. C + VEH= Control + Vehicle; M + VEH= Malaria + Vehicle; M +TH5487= Malaria + TH5487; M + O8= Malaria + O8. Data are mean ± SEM, (n=6), * P< 0.51, ** P< 0.01

Article Snippet: The TH5487 (catalogue no. 6749) and O8-OGG1 (catalogue no. 6236) OGG1 inhibitors were purchased from Tocris Bioscience, USA.

Techniques: Activity Assay, Concentration Assay, Control

Journal: Iranian Journal of Parasitology

Article Title: Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice

doi: 10.18502/ijpa.v19i4.17163

Figure Lengend Snippet: Effect of modulating OGG1 activity on serum IL-4 concentration on day 5 post P. berghei inoculation. C + VEH= Control + Vehicle; M + VEH= Malaria + Vehicle; M +TH5487= Malaria + TH5487; M + O8= Malaria + O8. Data are mean ± SEM, (n=6)

Article Snippet: The TH5487 (catalogue no. 6749) and O8-OGG1 (catalogue no. 6236) OGG1 inhibitors were purchased from Tocris Bioscience, USA.

Techniques: Activity Assay, Concentration Assay, Control

Journal: Iranian Journal of Parasitology

Article Title: Modulation of 8-Oxoguanine DNA Glycosylase 1 (OGG1) Alleviated Anemia Severity and Excessive Cytokines Release during Plasmodium berghei Malaria in Mice

doi: 10.18502/ijpa.v19i4.17163

Figure Lengend Snippet: Effect of modulating OGG1 activity on serum IL-17A concentration on day 5 post P. berghei inoculation. C + VEH= Control + Vehicle; M + VEH= Malaria + Vehicle; M +TH5487= Malaria + TH5487; M + O8= Malaria + O8. Data are mean ± SEM, (n=6)

Article Snippet: The TH5487 (catalogue no. 6749) and O8-OGG1 (catalogue no. 6236) OGG1 inhibitors were purchased from Tocris Bioscience, USA.

Techniques: Activity Assay, Concentration Assay, Control

Journal: Journal of Extracellular Vesicles

Article Title: The 8‐oxoguanine DNA glycosylase‐synaptotagmin 7 pathway increases extracellular vesicle release and promotes tumour metastasis during oxidative stress

doi: 10.1002/jev2.12505

Figure Lengend Snippet: SYT7 is a potential target gene of OGG1. (A, B) After A549 cells were transfected with scrambled or OGG1 siRNA for 48 h, the mRNA levels of OGG1 (A) and SYT7 (B) were examined. (C) Cells were treated as panel (A). Western blotting was used to determine the protein levels of OGG1 and SYT7. Right, quantification of OGG1 and SYT7. (D, E) The mRNA (D) and protein (E) levels of SYT7 in the WT and OGG1‐KO cells were examined by RT‐qPCR and Western blotting. (F) Analysis of CpG islands 2 kb upstream of the TSS region of SYT7. Upper graph: A plot of the ratio of observed GC content to expected GC content. Middle graph: GC content percentage. Lower graph: The CpG island was predicted to exist at −844 to −55 nucleotides (nt) upstream of the TSS region of SYT7. (G, H) The mRNA (G) and protein (H) levels of SYT7 in A549 cells treated with or without 20 µM H 2 O 2 were analysed. Right, quantification of SYT7 level. (I, J) After cells were treated with 20 µM H 2 O 2 and 10 mM NAC for 24 h, the mRNA (I) and protein (J) levels of SYT7 were analysed. Right, quantification of SYT7. (K, L) After cells were treated with or without 10 µM Th5487 for 24 h, the mRNA (K) and protein (L) levels of SYT7 were analysed. Right, quantification of SYT7 level. (M, N) After cells were treated with or without 10 µM OGG1‐IN‐08 for 24 h, the mRNA (M) and protein (N) levels of SYT7 were analysed. Right, quantification of SYT7 level. All of the data are expressed as mean values ± SEM ( n = 3); ** p < 0.01, *** p < 0.001. (Student's t ‐test).

Article Snippet: In the Th5487 treatment experiment, after 10 days of xenograft, BALB/c‐Nude mice (GemPharmatech Co., Ltd., Nanjing, China) received daily intraperitoneal injections of Th5487 (30 mg/kg, 200 µL) daily.

Techniques: Transfection, Western Blot, Quantitative RT-PCR

Journal: Journal of Extracellular Vesicles

Article Title: The 8‐oxoguanine DNA glycosylase‐synaptotagmin 7 pathway increases extracellular vesicle release and promotes tumour metastasis during oxidative stress

doi: 10.1002/jev2.12505

Figure Lengend Snippet: OGG1 promotes the binding of NF‐κB to target sites on the SYT7 promoter. (A) Analysis of the SYT7 promoter. NF‐κB binding sites in the SYT7 promoter region were predicted by JASPAR database. (B) The NF‐κB level in the cytoplasm and nucleus was analysed by Western blotting. Right, quantification of NF‐κB level. (C, D) A549 cells were exposed to 20 ng/mL TNF‐α for 4 h, and the mRNA (C) and protein (D) levels were analysed. Right, quantification of SYT7 level. (E, F) Cells were treated with or without 10 µM JSH‐23, and the mRNA (E) and protein (F) levels of SYT7 were analysed. Right, quantification of SYT7 level. (G, H) Cells were treated with or without 10 µM JSH‐23 or/and 10 µM OGG1‐IN‐08. The mRNA (G) and protein (H) levels of SYT7 were analysed. Right, quantification of SYT7 level. (I, J) 8‐oxoG increased the occupancy of OGG1 (I) or NF‐κB (J) on the SYT7 promoter. Purified OGG1 protein or nucleoprotein was subjected to EMSA with different FAM‐labelled probes. The arrow indicated the position of the NF‐κB or OGG1‐probe complex. (K) Occupancy of NF‐κB on 8‐oxoG‐containing DNA in nuclear extracts. Nuclear extracts (50 µg per sample) were incubated with G or 8‐oxoG probes for 12 h, and the protein‐DNA complexes were analysed by Western blotting. (L, M) ChIP‐qPCR assay determined the H 2 O 2 ‐induced enrichment of OGG1 (L) or NF‐κB (M) on the SYT7 promoter. (N) ChIP‐qPCR detected the enrichment of NF‐κB at the SYT7 promoter in WT or OGG1‐KO cells under oxidative stress. (O) EMSA detected the effect of OGG1‐IN‐08 and Th5487 exposure on OGG1 binding to oxidised DNA strands. 10 µM OGG1‐IN‐08 or 10 µM Th5487 was incubated with 100 ng OGG1. (P) ChIP‐qPCR analysis of NF‐κB enrichment at the SYT7 promoter under oxidative stress with or without Th5487 treatment. (Q) ChIP‐qPCR analysis of NF‐κB enrichment at the SYT7 promoter under oxidative stress with or without OGG1‐IN‐08 treatment. All of the data are expressed as mean values ± SEM ( n = 3); * p < 0.05, ** p < 0.01, *** p < 0.001. (Student's t ‐test).

Article Snippet: In the Th5487 treatment experiment, after 10 days of xenograft, BALB/c‐Nude mice (GemPharmatech Co., Ltd., Nanjing, China) received daily intraperitoneal injections of Th5487 (30 mg/kg, 200 µL) daily.

Techniques: Binding Assay, Western Blot, Purification, Incubation

Journal: Journal of Extracellular Vesicles

Article Title: The 8‐oxoguanine DNA glycosylase‐synaptotagmin 7 pathway increases extracellular vesicle release and promotes tumour metastasis during oxidative stress

doi: 10.1002/jev2.12505

Figure Lengend Snippet: Overexpression of SYT7 partially restores the down‐regulation of migration caused by OGG1 deletion. (A) After overexpressing SYT7 in cells knocked down OGG1, transwell assay was used to detect migrated cells. Right, quantification of migrated cells. (B) Cells were treated as panel (A). Protein levels of EMT markers were verified by western blot assays. Right, quantitative analysis. (C) After overexpressing SYT7 in cells knocked out OGG1, transwell assay was used to detect migrated cells. Right, quantification of migrated cells. (D) Cells were treated as panel (C). Protein levels of EMT markers were verified by western blot assays. Right, quantitative analysis. (E) The total protein concentration of isolated EVs was determined by BCA assay. (F) Western blotting analysed equal amounts of cell‐isolated EVs for markers content. Right, quantitative analysis of markers. (G) The content of E‐cadherin in equal amounts of EVs was quantified by Western blotting. (H) Transwell assay detected the invasion and migration ability of A549 cells after Th5487 exposure. Right, quantification of migrated cells. (I) Cells were treated with Th5487 for 24 h. EMT‐related markers were detected by Western blotting. Right, quantification of the marks. (J) After cells were treated with Th5487 for 24 h, EVs from the culture supernatant were isolated by ultra‐fractionation. The total protein concentration of isolated EVs was determined by BCA assay. (K) ELISA determined the content of CD63 in isolated EVs. (L) Western blotting analysed equal amounts of cell‐isolated EVs for markers content. Right, quantitative analysis of markers. (M) The content of E‐cadherin in equal amounts of EVs was quantified by Western blotting. All of the data are expressed as mean values ± SEM ( n = 3); ** p < 0.01, *** p < 0.001. (Student's t ‐test).

Article Snippet: In the Th5487 treatment experiment, after 10 days of xenograft, BALB/c‐Nude mice (GemPharmatech Co., Ltd., Nanjing, China) received daily intraperitoneal injections of Th5487 (30 mg/kg, 200 µL) daily.

Techniques: Over Expression, Migration, Transwell Assay, Western Blot, Protein Concentration, Isolation, BIA-KA, Fractionation, Enzyme-linked Immunosorbent Assay

Journal: Journal of Extracellular Vesicles

Article Title: The 8‐oxoguanine DNA glycosylase‐synaptotagmin 7 pathway increases extracellular vesicle release and promotes tumour metastasis during oxidative stress

doi: 10.1002/jev2.12505

Figure Lengend Snippet: OGG1 deletion reduces lung cancer metastasis in vivo. (A) Schematic diagram of the experimental process of zebrafish xenograft. CD‐Dil‐labelled A549 cells (red) were injected into the perivitelline space of each zebrafish. (B) A zebrafish xenograft model was used to evaluate the metastatic ability of A549 cells after OGG1 knockout. Representative images of zebrafish were shown. (C) Schematic diagram of establishing a lung cancer metastasis model by injecting A549 cells into the tail vein. (D) In vivo bioluminescence imaging of mice 40 days after cell injection. (E) Gross anatomy of mouse lungs. (F) Representative HE staining images of lungs from each group. Scale bar: 200 µm. (G, H) Zebrafish juvenile survival rate (G) and heart rate (H) at 120 hpf. Conventional one‐way ANOVA test was performed considering the experimental group as the independent variable. ns stands for no significance. (I) Schematic diagram of an experimental zebrafish xenograft experiment using A549 cells to test the anti‐tumour effect of Th5487. (J) Zebrafish xenograft model was used to evaluate the effect of Th5487 on A549 cell metastasis. Representative images of zebrafish were shown. (K) A mouse lung metastasis model was established by injecting A549 cells into the tail vein. Ten days after cell injection, mice were intraperitoneally injected with 30 mg/kg Th5487 or a corresponding volume of PBS every 2 days. (L) Gross anatomy of mouse liver. (M) Representative HE staining images of lungs and liver in each group. Scale bar: 50 µm. (N) Representative images from IHC staining for SYT7. Scale bar: 50 µm. All of the data except zebrafish juvenile survival rate and heart rate are expressed as mean values ± SEM ( n = 3); ** p < 0.01, *** p < 0.001. (Student's t ‐test).

Article Snippet: In the Th5487 treatment experiment, after 10 days of xenograft, BALB/c‐Nude mice (GemPharmatech Co., Ltd., Nanjing, China) received daily intraperitoneal injections of Th5487 (30 mg/kg, 200 µL) daily.

Techniques: In Vivo, Injection, Knock-Out, Imaging, Staining, Immunohistochemistry