Journal:

Article Title: Oxidative DNA damage and its repair in rat spleen following subchronic exposure to aniline

doi: 10.1016/j.taap.2008.08.010

Figure Lengend Snippet: BER activity in the splenic NEs of control and aniline-treated rats. The assay was conducted with end-labeled oligonucleotides (8-oxoG:C) which are targeted by OGG1 (see Methods for details). (A) Autoradiogram of incision products (P) generated over time by cleavage of end-labeled double-stranded oligonucleotides carrying the *8-oxoG:C adducts (S). Negative control (without NEs) and positive control (recombinant OGG1) are included in external lanes. (B) Values from Phosphorimager quantitation of products, generated with NEs from 6 rats per group and 3 cleavage assays per extract, were averaged and plotted as means ± SD. * Indicates that the cleavage is significantly different from controls for each respective time point (p < 0.05).

Article Snippet: The membrane was then blocked with 10% non-fat dry milk and incubated overnight at 4 °C with the rabbit anti-OGG1 antibody (Novus Biologicals, Littleton, CO) at the final concentration of 1.8 μg/ml (1:600 dilution).

Techniques: Activity Assay, Control, Labeling, Generated, Negative Control, Positive Control, Recombinant, Quantitation Assay

Journal:

Article Title: Oxidative DNA damage and its repair in rat spleen following subchronic exposure to aniline

doi: 10.1016/j.taap.2008.08.010

Figure Lengend Snippet: BER activity in the MEs of spleens from control and aniline-treated rats. The assay was conducted with end-labeled oligonucleotides (8-oxoG:C) which are targeted by OGG1 (see Methods for details). (A) Autoradiogram of incision products (P) generated over time by cleavage of end-labeled double-stranded oligonucleotides carrying the *8-oxoG:C adducts (S). Negative control (without MEs) and positive control (recombinant OGG1) are included in external lanes. (B) Values from Phosphorimager quantitation of products, generated with MEs from 6 rats per group and 3 cleavage assays per extract, were averaged and plotted as means ± SD. * Indicates that the cleavage is significantly different from controls for each respective time point (p < 0.05).

Article Snippet: The membrane was then blocked with 10% non-fat dry milk and incubated overnight at 4 °C with the rabbit anti-OGG1 antibody (Novus Biologicals, Littleton, CO) at the final concentration of 1.8 μg/ml (1:600 dilution).

Techniques: Activity Assay, Control, Labeling, Generated, Negative Control, Positive Control, Recombinant, Quantitation Assay

Journal:

Article Title: Oxidative DNA damage and its repair in rat spleen following subchronic exposure to aniline

doi: 10.1016/j.taap.2008.08.010

Figure Lengend Snippet: Real-time PCR analysis of OGG1 gene expression in the spleens of control and aniline-treated rats. Total RNA was extracted from spleen tissues, real-time PCR was performed, and the fold change in mRNA expression was determined. Values are means ± SD. *p< 0.05.

Article Snippet: The membrane was then blocked with 10% non-fat dry milk and incubated overnight at 4 °C with the rabbit anti-OGG1 antibody (Novus Biologicals, Littleton, CO) at the final concentration of 1.8 μg/ml (1:600 dilution).

Techniques: Real-time Polymerase Chain Reaction, Gene Expression, Control, Expressing

Journal:

Article Title: Oxidative DNA damage and its repair in rat spleen following subchronic exposure to aniline

doi: 10.1016/j.taap.2008.08.010

Figure Lengend Snippet: Western blot detection of OGG1 in the NEs (A) and MEs (C) from control and aniline-treated rats. Lanes 1-3: controls; lanes 4-6: aniline-treated. (B, D) Densitometric analyses of OGG1 bands from control and aniline-treated rats. The densitometric analysis of the protein bands was done using Eagle Eye II software. Data from aniline-treated spleen samples are presented in comparison to untreated controls, which were set at 100. Values are means ± SD of three determinations. *p< 0.05.

Article Snippet: The membrane was then blocked with 10% non-fat dry milk and incubated overnight at 4 °C with the rabbit anti-OGG1 antibody (Novus Biologicals, Littleton, CO) at the final concentration of 1.8 μg/ml (1:600 dilution).

Techniques: Western Blot, Control, Software, Comparison

Journal:

Article Title: Oxidative DNA damage and its repair in rat spleen following subchronic exposure to aniline

doi: 10.1016/j.taap.2008.08.010

Figure Lengend Snippet: Immunohistochemistry of OGG1 in the spleen of control and aniline-treated rats. A: control spleen; B: aniline-treated spleen. Controls showed scattered immunoreactivity for OGG1, whereas aniline-treated spleens showed strong immunoreactivity for OGG1 confined to the red pulp areas of the spleen (see Methods for details of immunohistochemistry).

Article Snippet: The membrane was then blocked with 10% non-fat dry milk and incubated overnight at 4 °C with the rabbit anti-OGG1 antibody (Novus Biologicals, Littleton, CO) at the final concentration of 1.8 μg/ml (1:600 dilution).

Techniques: Immunohistochemistry, Control

Journal: Journal of Biological Chemistry

Article Title: Myostatin Induces DNA Damage in Skeletal Muscle of Streptozotocin-induced Type 1 Diabetic Mice

doi: 10.1074/jbc.m113.483115

Figure Lengend Snippet: FIGURE 3. Absence of Mstn abrogated STZ-induced changes in p63 and REDD1 signaling. A, Western blot analysis (i) and densitometric analysis (ii) of p-Akt1/2/3,Akt1/2/3,IRS-1andIGF-1inWT-C(lane1),WT-STZ(lane2),Mstn/-C(lane3)andMstn/-STZ(lane4)Gastrocnemiusmuscleproteinlysates.*,p 0.05, **, p 0.01 when compared with WT-C; ˆˆ, p 0.01 when compared with Mstn/-C. GAPDH was used as an internal control for equal protein loading on the gel (n 7). B, Western blot analysis (i) and densitometric analysis (ii) of REDD1, OGG1, p53, and p63 protein levels in WT-C (lane 1), WT-STZ (lane 2), Mstn/-C (lane 3), and Mstn/-STZ (lane 4) Gastrocnemius muscle. GAPDH was used as an internal control for equal protein loading on the gel (n 5). *, p 0.05, ***, p 0.001 when compared with WT-C. Immunohistochemistry for REDD1 (C) and OGG1 (D) was performed on cryosections of Tibialis anterior muscles from WT-C and WT-STZ (i) and Mstn/-C and Mstn/-STZ (ii) mice. The fluorescence was viewed under a Leica upright microscope and images were taken at 5 magnification. Increased or decreased fluorescence (green) indicates changes in expression of REDD1 (C) or OGG1 (D); scale bar represents 100 m (n 3). E, ROS production was measured in proliferating C2C12 myoblasts treated with STZ1 for 48 h in Permanox chamber slides using the CM-H2DCFDA fluorescent probe. The fluorescence was viewed under a Leica upright microscope and images were taken at 10 magnification. Increased fluorescence (green) intensity is directly proportional to increased ROS production; scale bar represents 100 m (n 2).

Article Snippet: The immunohistochemistry protocol for REDD1 or OGG1 primary antibodies was followed as per manufacturer’s instructions (Proteintech).

Techniques: Western Blot, Control, Immunohistochemistry, Muscles, Fluorescence, Microscopy, Expressing

Journal: Journal of Biological Chemistry

Article Title: Myostatin Induces DNA Damage in Skeletal Muscle of Streptozotocin-induced Type 1 Diabetic Mice

doi: 10.1074/jbc.m113.483115

Figure Lengend Snippet: FIGURE 4. STZ-induced Mstn signaling in vitro via Foxa2 leads to oxidative stress-induced DNA damage, which was attenuated by Ant1. A, represent- ative Western blot (i) and densitometric analysis (ii) showing Foxa2 levels in protein lysates obtained from proliferating C2C12 cells treated with STZ1 for 48 h (lane 1-Untreated; lane 2-STZ1 treated). GAPDH was used as an internal control for equal protein loading on the gel (*, p 0.05; n 3). B, (i) Western blot and (ii) densitometric analysis showing protein levels of Mstn, p-Smad2/3, Smad2/3, p-Akt1/2/3, and Akt1/2/3 in proliferating C2C12 cells untreated (lane 1), treated with STZ1 (lane 2) or STZ2 (lane 3) or Ant1 (lane 4) for 48 h, pretreated with Ant1 for 1 h followed by either STZ1 (lane 5) or STZ2 (lane 6) treatment for 48 h. GAPDH was used as an internal control for equal protein loading on the gel (n 3) (*, p 0.05, **, p 0.01 when compared with untreated cells; ˆ, p 0.05 when compared with Ant1-treated cells). Western blot analysis (i) and densitometric analysis (ii) of p63, REDD1, and OGG1 in protein lysates obtained from proliferating C2C12 cells transfected with either (C) p-FLAG-CMV2 or p-FLAG-Foxa2 or (D) scrambled -ve control siRNA or Foxa2-siRNA. (Lanes 1 and 2-untreated; lanes 3 and 4-STZ1 treated; lanes 5 and 6-STZ2 treated). GAPDH was used as an internal control for equal protein loading on the gel (*, p 0.05). (n 3).

Article Snippet: The immunohistochemistry protocol for REDD1 or OGG1 primary antibodies was followed as per manufacturer’s instructions (Proteintech).

Techniques: In Vitro, Western Blot, Control, Transfection

Journal: Oncogene

Article Title: Synergistic lethality between PARP-trapping and alantolactone-induced oxidative DNA damage in homologous recombination-proficient cancer cells

doi: 10.1038/s41388-020-1191-x

Figure Lengend Snippet: a Both hydrogen peroxide and auranofin sensitize cancer cells to olaparib. PC-3 cells were treated by 1, 2, 4, 8, or 16 μM olaparib alone or combined with 100 μM H 2 O 2 or 0.5 μM auranofin for 72 h. Olaparib in combination with 100 μM H 2 O 2 or 0.5 μM auranofin dose-dependently inhibited the growth of PC-3 cells, while olaparib alone had no impact. NAC blocked H 2 O 2 or auranofin-induced olaparib sensitization. b CI values between auranofin and olaparib indicate strong synergy. PC-3 or A549 cells were treated by 0.5 μM auranofin and the indicated concentrations of olaparib for 72 h. c Western blot verification of shRNA-mediated knockdown of OGG1 and PARP1 in PC-3 cells. d MTT proliferation assay. Wild-type, OGG1-depleted or inhibited by 8 μM O8, and PARP1-depleted PC-3 cells were treated by 2, 4, 8, or 16 μM olaparib in combination with 10 μM ATL for 72 h. e MTT assay. Wild-type or PARP1 depleted PC-3 cells were treated by 10 μM ATL or the combination of 10 μM ATL and 10 μM olaparib (Ola) for 72 h. PARP1 knockdown did not sensitize PC-3 cells to 10 μM ATL. f Detection of chromatin-bound PARP1 by Western blot. PC-3 cells were treated by 10 μM ATL, 10 μM olaparib (Ola), 10 μM veliparib (Vel), or the combination of 10 μM ATL with 10 μM olaparib or veliparib for 24 h. n.s. not significant, *** p < 0.001 vs. vehicle control.

Article Snippet: Primary antibodies include γH2AX-pS139 polyclonal antibody (ab11174), poly(ADP-ribose) polymer (ab14459), and BrdU (ab6326, ab8152) (Abcam); cleaved caspase-3 (9664S), H3-pS10 (9706L), CHK1-pS317 (12302S), and CHK2-pT68 (2661S) (Cell Signaling, Danvers, MA, USA); RPA32 (bs-4182R), CHK1 (bs1681R), OGG1 (bs3687R), GAPDH (bs10900R), and β-actin (bsm33036M) (Bioss, Beijing, China); H3 (abs131869), H2AX (abs131731), CHK2 (abs131635) (Absin Bioscience, Shanghai, China); RPA32-pS4/8 (NBP1-23017) (NOVUS, Centennial, CO, USA), 53BP1 (A300-272A) (Bethyl, Montgomery, TX, USA), γH2AX-pS139 monoclonal antibody (14-9865-82) (ThermoFisher, Waltham, MA, USA).

Techniques: Western Blot, shRNA, Proliferation Assay, MTT Assay