primary antibodies against sirt3, p53, p21, atm, cyclind1, hmgb1, ogg1, p16, nf-κb, gapdh  (Proteintech)

Journal: bioRxiv

Article Title: Metformin prevented high glucose-induced endothelial reactive oxygen species via OGG1 in an AMPKα-Lin-28 dependent pathway

doi: 10.1101/2020.11.18.388462

Figure Lengend Snippet: High glucose decreased OGG1 expression while increased endothelial reactive oxygen species levels. (A-C) OGG1 mRNA and protein levels were tested after HG exposure for indicated periods. (B) is the statistical result of (A). (D) The expression of OGG1 in the aorta of diabetic mice were assessed via Immunofluorescence. (E-F) HUVECs were digested and incubated with DHE probe (1 μM) for 30 minutes after high glucose exposure for 12 hours. The mean intensities were obtained via flow Jo software. Scar bar =25μm. *P<0.05 versus NG group (mannitol control group). Values are the means±SD with N = 3.

Article Snippet: Primary antibodies anti-OGG1 (NB100-106, Novus Biologicals), anti-Lin28 ser 200 (20607, CST), anti-Lin28 (3695, CST), anti-AMPK (5831, CST), anti-Phospho-AMPKα (50081, CST), and ß-Actin (4970, CST) were obtained commercially.

Techniques: Expressing, Immunofluorescence, Incubation, Software, Control

Journal: bioRxiv

Article Title: Metformin prevented high glucose-induced endothelial reactive oxygen species via OGG1 in an AMPKα-Lin-28 dependent pathway

doi: 10.1101/2020.11.18.388462

Figure Lengend Snippet: Metformin decreased ROS and activated OGG1 in the HUVECs. (A-B) Endothelial ROS was measured by DHE probe after high glucose exposure with metformin or not. (B) is the stastical result of (A). (C-E) OGG1 mRNA and protein levels were tested after metformin (0.5 mM) exposure for 24 hours. (E) is the statistical result of (D). *P<0.05 versus NG group (mannitol control group). #P<0.05 versus HG group. Values are the means±SD with N = 3. (F-G) HUVECs were digested and incubated with DHE probe (1 μM) for 30 minutes after silencing OGG1 or not in the presence with HG and metformin. The mean intensities were obtained via flow Jo software. Scar bar =25μm. *P<0.05 versus HG plus metformin group. Values are the means±SD with N = 3.

Article Snippet: Primary antibodies anti-OGG1 (NB100-106, Novus Biologicals), anti-Lin28 ser 200 (20607, CST), anti-Lin28 (3695, CST), anti-AMPK (5831, CST), anti-Phospho-AMPKα (50081, CST), and ß-Actin (4970, CST) were obtained commercially.

Techniques: Control, Incubation, Software

Journal: bioRxiv

Article Title: Metformin prevented high glucose-induced endothelial reactive oxygen species via OGG1 in an AMPKα-Lin-28 dependent pathway

doi: 10.1101/2020.11.18.388462

Figure Lengend Snippet: The OGG1 expression stimulated by metformin was dependent on its mRNA stability in the HUVECs. (A) HUVECs were incubated with actinomycin D (Act D, 10 μg/ml) for 0, 6, and 12 hours, and then OGG1 mRNA was measured by qPCR. (B-C) OGG1 protein levels were tested by WB after CHX-treated HUVECs for 0, 6, and 12 hours. *P<0.05 versus control group (mannitol control group). Values are the means±SD with N = 3.

Article Snippet: Primary antibodies anti-OGG1 (NB100-106, Novus Biologicals), anti-Lin28 ser 200 (20607, CST), anti-Lin28 (3695, CST), anti-AMPK (5831, CST), anti-Phospho-AMPKα (50081, CST), and ß-Actin (4970, CST) were obtained commercially.

Techniques: Expressing, Incubation, Control

Journal: bioRxiv

Article Title: Metformin prevented high glucose-induced endothelial reactive oxygen species via OGG1 in an AMPKα-Lin-28 dependent pathway

doi: 10.1101/2020.11.18.388462

Figure Lengend Snippet: The mRNA stability of OGG1 triggered by metformin was dependent on AMPKα. (A-B) The phosphorylation of AMPKα and total AMPKα were measured after metformin-treated HUVECs. (C-E) OGG1 protein and mRNA levels were tested by WB and qPCR after metformin and high glucose treatment simultaneously with silencing of AMPKα or not in the HUVECs. (F-G) HUVECs were digested and incubated with DHE probe (1 μM) for 30 minutes after silencing AMPKα or not in the presence with HG and metformin. The mean intensities were obtained via flow Jo software. Scar bar =25μm. *P<0.05 versus control group. #P<0.05 versus HG plus metformin group. Values are the means±SD with N = 3.

Article Snippet: Primary antibodies anti-OGG1 (NB100-106, Novus Biologicals), anti-Lin28 ser 200 (20607, CST), anti-Lin28 (3695, CST), anti-AMPK (5831, CST), anti-Phospho-AMPKα (50081, CST), and ß-Actin (4970, CST) were obtained commercially.

Techniques: Phospho-proteomics, Incubation, Software, Control

Journal: bioRxiv

Article Title: Metformin prevented high glucose-induced endothelial reactive oxygen species via OGG1 in an AMPKα-Lin-28 dependent pathway

doi: 10.1101/2020.11.18.388462

Figure Lengend Snippet: Lin28 meditated OGG mRNA stability of OGG1 triggered by metformin. (A-B) The subcellular localization of Lin28 were measured after metformin-treated HUVECs in the presence of high glucose. (C-E) OGG1 protein and mRNA levels were tested by WB and qPCR after metformin and high glucose treatment simultaneously with silencing of Lin28 or not in the HUVECs. (F) The half-time hours of OGG1 mRNA was measured after silencing Lin-28 or not with actinomycin D (Act D, 10 μg/ml). *P<0.05 versus control group. Values are the means±SD with N = 3.

Article Snippet: Primary antibodies anti-OGG1 (NB100-106, Novus Biologicals), anti-Lin28 ser 200 (20607, CST), anti-Lin28 (3695, CST), anti-AMPK (5831, CST), anti-Phospho-AMPKα (50081, CST), and ß-Actin (4970, CST) were obtained commercially.

Techniques: Control

Journal: bioRxiv

Article Title: Metformin prevented high glucose-induced endothelial reactive oxygen species via OGG1 in an AMPKα-Lin-28 dependent pathway

doi: 10.1101/2020.11.18.388462

Figure Lengend Snippet: OGG1 expression induced by metformin was dependent on AMPKα phosphorylation of Lin-28. (A-B) The subcellular localization of Lin28 were measured after AMPKα silence or inhibitor pretreatment. *P<0.05 versus HG group. #P<0.05 versus HG plus metformin group. (C-D) OGG1 protein and mRNA levels were tested after silence Lin28 in the HUVECs with AMPKα plasmid or AICAR. *P<0.05 versus HG group. #P<0.05 versus HG plus ACICAR group. Values are the means±SD with N = 3.

Article Snippet: Primary antibodies anti-OGG1 (NB100-106, Novus Biologicals), anti-Lin28 ser 200 (20607, CST), anti-Lin28 (3695, CST), anti-AMPK (5831, CST), anti-Phospho-AMPKα (50081, CST), and ß-Actin (4970, CST) were obtained commercially.

Techniques: Expressing, Phospho-proteomics, Plasmid Preparation

Journal: bioRxiv

Article Title: Metformin prevented high glucose-induced endothelial reactive oxygen species via OGG1 in an AMPKα-Lin-28 dependent pathway

doi: 10.1101/2020.11.18.388462

Figure Lengend Snippet: The protective role of metformin on endothelial ROS was dependent on AMPKα/Lin-28/OGG1 pathway. (A-B) Endothelial ROS levels were assessed by DHE after Compound C or silence of Lin-28-treated HUVECs. *P<0.05 versus HG group. #P<0.05 versus HG plus metformin (Met) group. Values are the means±SD with N = 3.

Article Snippet: Primary antibodies anti-OGG1 (NB100-106, Novus Biologicals), anti-Lin28 ser 200 (20607, CST), anti-Lin28 (3695, CST), anti-AMPK (5831, CST), anti-Phospho-AMPKα (50081, CST), and ß-Actin (4970, CST) were obtained commercially.

Techniques:

Journal: bioRxiv

Article Title: Metformin prevented high glucose-induced endothelial reactive oxygen species via OGG1 in an AMPKα-Lin-28 dependent pathway

doi: 10.1101/2020.11.18.388462

Figure Lengend Snippet: OGG1 regulated HG-induced endothelial ROS via NFκB/NOX4 pathway. (A-B) NADPH oxidases were measured after HUVECs exposure to high glucose treatment with overexpression of OGG1. (B) was statistical result of (A). (C-D) NFκB activity was measured after OGG1 overexpression with high glucose treatment for 24 hours. (E-F) NOX4 protein level was assessed after NFκB silence with high glucose incubation. *P<0.05 versus Ctrl group. #P<0.05 versus HG group. NS P means no difference. Values are the means±SD with N = 3.

Article Snippet: Primary antibodies anti-OGG1 (NB100-106, Novus Biologicals), anti-Lin28 ser 200 (20607, CST), anti-Lin28 (3695, CST), anti-AMPK (5831, CST), anti-Phospho-AMPKα (50081, CST), and ß-Actin (4970, CST) were obtained commercially.

Techniques: Over Expression, Activity Assay, Incubation

Journal: Molecular Medicine Reports

Article Title: Association between oxidative DNA damage and the expression of 8-oxoguanine DNA glycosylase 1 in lung epithelial cells of neonatal rats exposed to hyperoxia

doi: 10.3892/mmr.2015.3339

Figure Lengend Snippet: Primers used for reverse transcription quantitative polymerase chain reaction.

Article Snippet: The sections were then blocked with 5% goat serum for 1 h at room temperature and incubated with goat polyclonal anti-OGG1 primary antibody (1:50; ab115841; Abcam) overnight at 4°C.

Techniques: Reverse Transcription

Journal: Molecular Medicine Reports

Article Title: Association between oxidative DNA damage and the expression of 8-oxoguanine DNA glycosylase 1 in lung epithelial cells of neonatal rats exposed to hyperoxia

doi: 10.3892/mmr.2015.3339

Figure Lengend Snippet: Expression of OGG1 (red stain) is predominantly localized in the cytoplasm on (A) day 1 and (B) day 5 in normoxia-exposed rats and on (C) day 1 in hyperoxia-exposed rats. On days (D) 3, (E) 5 and (F) 7 in hyperoxia (magnification, ×400), the expression of OGG1 increased in the cytoplasm and the nucleus. (G and H) Western blotting revealed similar patterns of expression. Data are expressed as the mean ± standard deviation ( * P<0.05, ** P<0.01 as compared with the normoxia group). OGG1, 8-oxoguanine DNA glycosylase 1; N, normoxia; H, hyperoxia.

Article Snippet: The sections were then blocked with 5% goat serum for 1 h at room temperature and incubated with goat polyclonal anti-OGG1 primary antibody (1:50; ab115841; Abcam) overnight at 4°C.

Techniques: Expressing, Staining, Western Blot, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Association between oxidative DNA damage and the expression of 8-oxoguanine DNA glycosylase 1 in lung epithelial cells of neonatal rats exposed to hyperoxia

doi: 10.3892/mmr.2015.3339

Figure Lengend Snippet: Protein expression of OGG1 in cultured neonatal rat alveolar epithelial type II cells. (A) Western blotting and (B) densitometric quantification of the protein expression of OGG1 following different durations of hyperoxia or normoxia exposure. Data are expressed as the mean ± standard deviation ( * P<0.05; ** P<0.01 as compared with the normoxia group). OGG1, 8-oxoguanine DNA glycosylase 1.

Article Snippet: The sections were then blocked with 5% goat serum for 1 h at room temperature and incubated with goat polyclonal anti-OGG1 primary antibody (1:50; ab115841; Abcam) overnight at 4°C.

Techniques: Expressing, Cell Culture, Western Blot, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Association between oxidative DNA damage and the expression of 8-oxoguanine DNA glycosylase 1 in lung epithelial cells of neonatal rats exposed to hyperoxia

doi: 10.3892/mmr.2015.3339

Figure Lengend Snippet: mRNA expression levels of OGG1 in lung tissues and AECII cells. (A) Neonatal rat lung tissues and (B) neonatal rat AECII cells exposed to hyperoxia or normoxia. Data are expressed as the mean ± standard deviation (P>0.05 for all comparisons). AECII, alveolar epithelial type II cells; OGG1, 8-oxoguanine DNA glycosylase 1.

Article Snippet: The sections were then blocked with 5% goat serum for 1 h at room temperature and incubated with goat polyclonal anti-OGG1 primary antibody (1:50; ab115841; Abcam) overnight at 4°C.

Techniques: Expressing, Standard Deviation