rabbit polyclonal anti ogg1 antibody  (Novus Biologicals)

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: Table 1 The sequences of the primers used for genotyping

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques:

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: Table 3 Sequence information of negative control siRNA (NC), OGG1 siRNA and p53 siRNA

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Sequencing, Negative Control

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: Table 4 The sequences of the primers used for qRT-PCR in this study

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques:

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: Silencing of OGG1 promotes and overexpressing OGG1 inhibits cell senescence in lung cells A549 and BEAS-2B cells were transfected with siRNA-NC or siOGG1 (VC or OGG1) for 24 h and then treated with 25 μg/mL BLM for 24 h. (A–D) Representative images of SA β-galactosidase activity staining. Scale bar: 200 μm. (E,F) Statistical analysis results of SA-β-gal-positive lung cells in (A) and (B). (G–J) The protein expression of OGG1, the cellular senescence marker protein p21 and the DNA damage-associated protein p-H2AX were detected by western blot analysis. Tubulin protein was used for endogenous normalization. * P<0.05 vs NC or VC; #P<0.05 vs NC+BLM or VC+BLM. Data are expressed as the mean±SD of the experiments performed in triplicate.

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Transfection, Activity Assay, Staining, Expressing, Marker, Western Blot

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: OGG1 modulates cell transformation in A549 cells in the presence of BLM A549 cells were transfected with siRNA-NC or siOGG1 (A,C), VC or OGG1 (B,D) 24 h and then treated with 25 μg/mL BLM for 24 h. EMT markers, including FN1, N-cadherin and α-SMA proteins, were detected by western blot analysis. *P<0.05 vs NC or VC; #P<0.05 vs NC+BLM or VC+BLM. Data are expressed as the mean±SD of the experiments performed in triplicate.

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Transformation Assay, Transfection, Western Blot

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: OGG1 modulates cell cycle transition in BLM-induced lung cells A549 cells were transfected with siRNA-NC or siOGG1 (VC or OGG1) for 24 h and then treated with 25 μg/mL BLM for 24 h. (A,C) The population of cells at the G1, S and G2/M phase was detected by flow cytometry. (B,D) The statistical analysis results of lung cell numbers in different stages of the cell cycle in (A) and (C). *P<0.05 vs NC or VC; # P<0.05 vs NC+BLM or VC+BLM. Data are expressed as the mean±SD of the experiments performed in triplicate.

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Transfection, Flow Cytometry

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: OGG1 regulates TERT and LaminB1 mRNA and protein expressions in BLM-induced A549 cells A549 cells were transfected with siRNA-NC or siOGG1 (VC or OGG1) for 24 h and then treated with 25 μg/mL BLM for 24 h. The expressions of TERT and LaminB1 mRNA (A,B) and proteins (C–F) in A549 cells was detected by qPCR and western blot analysis, respectively. Tubulin protein was used for endogenous normalization. *P<0.05 vs NC or VC; #P<0.05 vs NC+BLM or VC+BLM. Data are expressed as the mean±SD of the experiments performed in triplicate.

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Transfection, Western Blot

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: OGG1 knockdown modulates senescence-related factors in A549 cells A549 cells were transfected with siRNA-NC or siOGG1 for 24 h and then treated with 25 μg/mL BLM for 24 h. The levels of SASP (A), including IL-1α, IL-1β, IL-6, IL-8, CXCL1/CXCL2, and MMP-3, reduced transcripts with senescence (B), including LBR, ANP32B, FAM129A, CBS, SSRP1, PARP1, and PTMA, increased transcripts with senescence (C), including LRP10, FILIP1L, ELMOD1, ARRDC4, TMEM30A, PDLIM1 and CCND3, were detected by qPCR assay. *P<0.05 vs NC; #P<0.05 vs NC+BLM. Data are expressed as the mean±SD of the experiments performed in triplicate.

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Knockdown, Transfection

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: OGG1 regulates p53 signaling in BLM-induced lung cells (A–D) A549 cells were transfected with siRNA-NC or siOGG1 (VC or OGG1) for 24 h and then treated with 25 μg/mL BLM for 24 h. The expressions of p-p53 and p53 protein in lung cells was detected by western blot analysis. (E) Immunofluorescence assay showing the colocalization of OGG1 with p53 in A549 cells. (F) Coimmunoprecipitation experiments showed the interaction of OGG1 with p53 in A549 cells. (G) Western blot analysis was used to determine the silencing effect of p53 siRNA. p53 siRNA was cotransfected with siRNA-NC or siOGG1 for 24 h and then treated with 25 μg/mL BLM for 24 h. (H,I) Representative images of SA β-galactosidase activity staining. Scale bar: 200 μm. (J,K) The expressions of OGG1, p21, p-H2AX, p-p53 and p53 proteins were detected by western blot analysis. Tubulin protein was used for endogenous normalization. *P<0.05 vs NC+BLM; #P<0.05 vs siOGG1+BLM. Data are expressed as the mean±SD of the experiments performed in triplicate.

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Transfection, Western Blot, Immunofluorescence, Activity Assay, Staining

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: Impairment in the DNA repair activity of OGG1 promotes lung cell senescence A549 cells were transfected with VC, OGG1, OGG1 K249Q (249), D268A (268), or R304 W (304) for 24 h and then treated with 25 μg/mL BLM for 24 h (A) Representative images of SA β-gal activity staining. (B) Statistical analysis results of SA-β-gal-positive lung cells in (A). (C,D) The expressions of OGG1, p21, p-H2AX, p-p53 and p53 in lung cells were detected by western blot analysis. Tubulin protein was used for endogenous normalization. *P<0.05 vs VC; #P<0.05 vs VC+BLM; & P<0.05 vs OGG1+BLM. Data are expressed as the mean±SD of the experiments performed in triplicate.

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Activity Assay, Transfection, Staining, Western Blot

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: Cell senescence is augmented in OGG1 -knockout mice (A) Genomic PCR of wild-type (WT) and OGG1 tail DNA: 429 bp OGG1 and 490 bp OGG1 (WT) amplification product. (B) The expressions of OGG1 protein in lung tissues from WT and OGG1-knockout mice were detected by western blot analysis to further verify the lack of OGG1 protein. WT and OGG1–/– mice were intratracheally treated with 50 μL of physiological saline (normal control) or BLM on day 0. Then, the mice were sacrificed at 21 days posttreatment. (C) The lung sections were stained with Masson. Scale bar: 300 μm. (D) The lung sections were stained with SA-β-gal. Scale bar: 300 μm .

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Knock-Out, Amplification, Western Blot, Saline, Control, Staining

Journal: Acta Biochimica et Biophysica Sinica

Article Title: 8-Oxoguanine DNA glycosylase protects cells from senescence via the p53-p21 pathway

doi: 10.3724/abbs.2023264

Figure Lengend Snippet: A schematic overview of the effects of OGG1 on cell senescence during pulmonary fibrosis OGG1 participates in two processes (normal repair and excessive repair) in IPF. Under normal physiological conditions, when lung epithelial cells and lung fibroblasts are damaged by BLM, OGG1 inhibits the p53-p21 signaling pathway by interacting with p53, inhibiting cell senescence and maintaining normal functions (normal repair). In the pathological environment of pulmonary fibrosis, when lung epithelial cells and lung fibroblasts are induced by BLM, OGG1 can perform normal repair functions and maintain normal cell functions (normal repair). However, due to the continuous damage to lung cells, more OGG1 is needed to promote the repair of damage, and the regulation of OGG1 expression is uncontrolled. Subsequently, abnormally upregulated OGG1 induces EMT in the anti-aging lung epithelial cells and abnormal activation in the anti-aging lung fibroblasts through TGF-β/Smads, ultimately leading to the occurrence of IPF.

Article Snippet: After being washed with PBS, the cells were fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 in PBS for 15 min. After incubation with blocking solution for 2 h at room temperature, the cells were incubated with rabbit polyclonal anti-OGG1 antibody (1:200; Novus Biologicals, Littleton, USA) and monoclonal mouse anti-p53 (1:200; Cell Signaling Technology, Danvers, USA) overnight at 4°C.

Techniques: Expressing, Activation Assay

Journal: Journal of molecular biology

Article Title: Human CST Stimulates Base Excision Repair to Prevent the Accumulation of Oxidative DNA Damage

doi: 10.1016/j.jmb.2024.168672

Figure Lengend Snippet: (A) Representative images of proximity ligation assay (PLA) performed in HeLa cells with antibodies to STN1 or Pol β alone or in combination with and without H 2 O 2 treatment. Red, PLA foci; blue, DAPI. (B) Violin plot of PLA foci per nucleus. Results are representative of three independent, biological experiments. Bold dashed line represents the median, and dashed lines represent the first and third quartiles. (C) Proximity ligation assay (PLA) performed in HeLa cells with antibodies to STN1, OGG1, or in combination (STN1 + OGG1). n = 3 independent, biological experiments. Bold dashed line: median, dashed lines: first and third quartiles. (D) HEK293T cell extracts containing individually overexpressed CST components (either FLAG-CST, Myc-STN1 or FLAG-TEN1) that were either untreated or treated with 200 μM of H 2 O 2 were used to immunoprecipitate APE1, Pol β, FEN1 or LIGI and immunobloted with either anti-FLAG (for CTC1 and TEN1) or anti-Myc for STN1. Lysates were pre-treated with benzonase prior to coimmunoprecipitation. Proteins are identified in the Western blot analysis. (**** p < 0.0001).

Article Snippet: Primary antibodies: 1:100 mouse α-STN1 (Novus, NBP2-01006), 1:100 rabbit α-POLB (Abbexa, abx304879), 1:100 rabbit α-PolA1 (Bethyl Laboratories, A302-850A), 1:100 rabbit α-XRCC1 (Genetex, GTX111712), 1:100 mouse α-XRCC1 (Invitrogen, MA1-12640), 1:100 rabbit α-PARP1 (Proteintech, 6520-1-Ig), 1:100 rabbit α-OGG1 (Proteintech, 15125-1-AP).

Techniques: Proximity Ligation Assay, Western Blot

Journal: Journal of molecular biology

Article Title: Human CST Stimulates Base Excision Repair to Prevent the Accumulation of Oxidative DNA Damage

doi: 10.1016/j.jmb.2024.168672

Figure Lengend Snippet: (A) OGG1 (2 nM, lanes 2, 4–15; 50 nM, lane 3) lesion base removal and phosphodiester bone cleavage activity were measured on a random sequence DNA substrate (5 nM) containing an 8-oxo-guanine (8-oxo-G) residue in the presence of increasing concentrations of CTC1/STN1/TEN1 (250, 750, 1500 nM) or CST (5, 15, 60 nM) as denoted in the figure. The red asterisk shown on the substrate in the figure represents the location of the 32 P radiolabel and a G* represents an 8-oxo-G residue. Gel shown is representative of at least three independent experiments. (B) Graph of the fold increase in OGG1 cleavage activity for the lanes containing the highest concentration of the CST complex and individual subunits. The random substrate (U1:T1) was generated by annealing primers in a 1:2 ratio. (**** p < 0.0001).

Article Snippet: Primary antibodies: 1:100 mouse α-STN1 (Novus, NBP2-01006), 1:100 rabbit α-POLB (Abbexa, abx304879), 1:100 rabbit α-PolA1 (Bethyl Laboratories, A302-850A), 1:100 rabbit α-XRCC1 (Genetex, GTX111712), 1:100 mouse α-XRCC1 (Invitrogen, MA1-12640), 1:100 rabbit α-PARP1 (Proteintech, 6520-1-Ig), 1:100 rabbit α-OGG1 (Proteintech, 15125-1-AP).

Techniques: Activity Assay, Sequencing, Residue, Concentration Assay, Generated

Journal: Journal of molecular biology

Article Title: Human CST Stimulates Base Excision Repair to Prevent the Accumulation of Oxidative DNA Damage.

doi: 10.1016/j.jmb.2024.168672

Figure Lengend Snippet: Figure 1. CST associates with BER proteins. (A) Representative images of proximity ligation assay (PLA) performed in HeLa cells with antibodies to STN1 or Pol b alone or in combination with and without H2O2 treatment. Red, PLA foci; blue, DAPI. (B) Violin plot of PLA foci per nucleus. Results are representative of three independent, biological experiments. Bold dashed line represents the median, and dashed lines represent the first and third quartiles. (C) Proximity ligation assay (PLA) performed in HeLa cells with antibodies to STN1, OGG1, or in combination (STN1 + OGG1). n = 3 independent, biological experiments. Bold dashed line: median, dashed lines: first and third quartiles. (D) HEK293T cell extracts containing individually overexpressed CST components (either FLAG- CST, Myc-STN1 or FLAG-TEN1) that were either untreated or treated with 200 mM of H2O2 were used to immunoprecipitate APE1, Pol b, FEN1 or LIGI and immunobloted with either anti-FLAG (for CTC1 and TEN1) or anti- Myc for STN1. Lysates were pre-treated with benzonase prior to coimmunoprecipitation. Proteins are identified in the Western blot analysis. (****p < 0.0001).

Article Snippet: Primary antibodies: 1:100 mouse a-STN1 (Novus, NBP2-01006), 1:100 rabbit a-POLB (Abbexa, abx304879), 1:100 rabbit a-PolA1 (Bethyl Laboratories, A302-850A), 1:100 rabbit a-XRCC1 (Genetex, GTX111712), 1:100 mouse a-XRCC1 (Invitrogen, MA1-12640), 1:100 rabbit a-PARP1 (Proteintech, 6520-1-Ig), 1:100 rabbit a-OGG1 (Proteintech, 15125-1-AP).

Techniques: Proximity Ligation Assay, Western Blot

Journal: Journal of molecular biology

Article Title: Human CST Stimulates Base Excision Repair to Prevent the Accumulation of Oxidative DNA Damage.

doi: 10.1016/j.jmb.2024.168672

Figure Lengend Snippet: Figure 2. The CST complex and individual subunits stimulate OGG1 activity on 8-oxo-G containing substrate. (A) OGG1 (2 nM, lanes 2, 4–15; 50 nM, lane 3) lesion base removal and phosphodiester bone cleavage activity were measured on a random sequence DNA substrate (5 nM) containing an 8-oxo-guanine (8-oxo-G) residue in the presence of increasing concentrations of CTC1/STN1/TEN1 (250, 750, 1500 nM) or CST (5, 15, 60 nM) as denoted in the figure. The red asterisk shown on the substrate in the figure represents the location of the 32P radiolabel and a G* represents an 8-oxo-G residue. Gel shown is representative of at least three independent experiments. (B) Graph of the fold increase in OGG1 cleavage activity for the lanes containing the highest concentration of the CST complex and individual subunits. The random substrate (U1:T1) was generated by annealing primers in a 1:2 ratio. (****p < 0.0001).

Article Snippet: Primary antibodies: 1:100 mouse a-STN1 (Novus, NBP2-01006), 1:100 rabbit a-POLB (Abbexa, abx304879), 1:100 rabbit a-PolA1 (Bethyl Laboratories, A302-850A), 1:100 rabbit a-XRCC1 (Genetex, GTX111712), 1:100 mouse a-XRCC1 (Invitrogen, MA1-12640), 1:100 rabbit a-PARP1 (Proteintech, 6520-1-Ig), 1:100 rabbit a-OGG1 (Proteintech, 15125-1-AP).

Techniques: Activity Assay, Sequencing, Residue, Concentration Assay, Generated