Journal: Molecular and Cellular Biology

Article Title: Poly(ADP-Ribose) Polymerase 1 Promotes Oxidative-Stress-Induced Liver Cell Death via Suppressing Farnesoid X Receptor ?

doi: 10.1128/MCB.00160-13

Figure Lengend Snippet: Oxidative stress-induced HepG2 cell death was attenuated by PARP1 inhibition. (A) HepG2 cells were exposed to H2O2 (3, 30, or 300 μM) for 12 h. Cells were trypsinized and were then counted in 3 fields by a hemocytometer with trypan blue dye (n = 3). Asterisks indicate significant differences (*, P < 0.05; **, P < 0.01) from the control group. (B) HepG2 cells were treated with H2O2 (3, 30, 300 or μM) for 12 h, and cellular PARP activity was assayed as described in Materials and Methods. Data are expressed as means ± SEM. Asterisks indicate significant differences (*, P < 0.05; **, P < 0.01) from the control group. (C) HepG2 cells were pretreated either with PJ34 (15 μM) for 24 h, with PARP1 siRNA (50 nM) for 48 h, or with GW4064 (1 μM) for 24 h and were then exposed to H2O2 (3, 30, or 300 μM) for 12 h. Cells were trypsinized and were counted in 3 fields by a hemocytometer with trypan blue dye (n = 3). Asterisks indicate significant differences (**, P < 0.01) from the control group. (D) HepG2 cells were transfected with either an empty vector (p3flag-CMV), wild type PARP1 (wt-PARP1), or the enzymatic mutant PARP1 (mut-PARP1) at 1 mg/liter for 48 h and were then exposed to H2O2 (3, 30, or 300 μM) for 12 h. Cells were trypsinized and were counted in 3 fields by a hemocytometer with trypan blue dye (n = 3). Asterisks indicate significant differences (*, P < 0.05) from the control group. (E) After transfection with FXR siRNA (50 nM) for 72 h, HepG2 cells were treated either with PJ34 (15 μM) for 24 h, with PARP1 siRNA (50 nM) for 48 h, or with GW4064 (1 μM) for 24 h and were then exposed to H2O2 (3, 30, or 300 μM) for 12 h. Cells were trypsinized and were counted in 3 fields by a hemocytometer with trypan blue dye (n = 3). Asterisks indicate significant differences (*, P < 0.05) from the unrelated siRNA group. The efficiencies of PARP1 siRNA, FXR siRNA, and wt- or mut-PARP1 transfection were determined by a real-time RT-PCR assay (shown in Fig. S2 in the supplemental material).

Article Snippet: PARP1 activity was assayed using the universal colorimetric PARP assay kit (Trevigen), based on the incorporation of biotinylated ADP-ribose into histone proteins.

Techniques: Inhibition, Control, Activity Assay, Transfection, Plasmid Preparation, Mutagenesis, Quantitative RT-PCR

Journal: Molecular and Cellular Biology

Article Title: Poly(ADP-Ribose) Polymerase 1 Promotes Oxidative-Stress-Induced Liver Cell Death via Suppressing Farnesoid X Receptor ?

doi: 10.1128/MCB.00160-13

Figure Lengend Snippet: Inhibition of PARP1 promoted the transcription of FXR-dependent hepatoprotective genes. (A and B) Real-time RT-PCR assays of BSEP, FGF19, Foxm1b, and SHP in HepG2 cells. After treatment with 1 μM GW4064 (A) or 15 μM PJ34 (B) for 12 h, cells were treated or not treated with H2O2 (300 μM; 12 h). Data are expressed as means ± SEM. Significant differences from the control group (**, P < 0.01) or from the H2O2 group (#, P < 0.05) are indicated. (C) Real-time RT-PCR assays of BSEP, FGF19, Foxm1b, and SHP in HepG2 cells transfected with either 50 nM PARP1 siRNA or 50 nM unrelated siRNA for 48 h, with or without H2O2 treatment (300 μM; 12 h). Data are expressed as means ± SEM. Significant differences from the control group (*, P < 0.05; **, P < 0.01) or from the H2O2 group (#, P < 0.05) are indicated. (D) Real-time RT-PCR assays of BSEP, FGF19, Foxm1b, and SHP in HepG2 cells transfected with either an empty vector (p3flag-CMV), wt-PARP1, or mut-PARP1 at 1 mg/liter for 48 h. Data are expressed as means ± SEM. Asterisks indicate significant differences (*, P < 0.05) from the control group. (E and F) Relative FXRE- or mutant (mut-FXRE)-driven luciferase reporter activity in HepG2 cells treated with 1 μM GW4064 (E) or 15 μM PJ34 (F) for 24 h, with or without H2O2 treatment (300 μM; 12 h). Data are expressed as means ± SEM. Significant differences from the control group (**, P < 0.01) or from the H2O2 group (#, P < 0.05) are indicated. (G) Relative FXRE- or mut-FXRE-driven luciferase reporter activity in HepG2 cells transfected with 50 nM PARP1 siRNA or 50 nM unrelated siRNA for 48 h, with or without H2O2 treatment (300 μM; 12 h). Data are expressed as means ± SEM. Significant differences from the control group (**, P < 0.01) or from the H2O2 group (#, P < 0.05) are indicated. (H) Relative FXRE- or mut-FXRE-driven luciferase reporter activity in HepG2 cells treated with an empty vector (p3flag-CMV), wt-PARP1, or mut-PARP1 at 1 mg/liter for 48 h. Data are expressed as means ± SEM. Asterisks indicate significant differences (**, P < 0.01) from the control group.

Article Snippet: PARP1 activity was assayed using the universal colorimetric PARP assay kit (Trevigen), based on the incorporation of biotinylated ADP-ribose into histone proteins.

Techniques: Inhibition, Quantitative RT-PCR, Control, Transfection, Plasmid Preparation, Mutagenesis, Luciferase, Activity Assay

Journal: Molecular and Cellular Biology

Article Title: Poly(ADP-Ribose) Polymerase 1 Promotes Oxidative-Stress-Induced Liver Cell Death via Suppressing Farnesoid X Receptor ?

doi: 10.1128/MCB.00160-13

Figure Lengend Snippet: PARP1 poly(ADP-ribosyl)ated the LBD of FXR. (A) Nuclear extracts from HepG2 cells were subjected to an immunoprecipitation assay with an anti-FXR antibody, followed by a Western blot assay using an anti-PAR antibody. Cells were treated with 15 μM PJ34 for 24 h in the presence or absence of H2O2 (300 μM; 12 h). (B) Nuclear extracts from HepG2 cells were subjected to an immunoprecipitation assay with an anti-FXR antibody, followed by a Western blot assay using an anti-PAR antibody. Cells were transfected with PARP1 siRNA or unrelated siRNA at 50 nM for 48 h with or without H2O2 treatment (300 μM; 12 h). (C) Recombinant FXR proteins were incubated either with a vehicle (PBS), with PARP1, NAD+, and active DNA, or with PARP1, NAD+, active DNA, and 3AB, as indicated. Western blot assays were used to detect the poly(ADP-ribosyl)ation levels of FXR. (D) (Top) Diagram of GST-tagged human FXR with its domains. (Center) Purified GST-FXR fragments are shown after Coomassie staining. (Bottom) Bacterially expressed GST-FXR deletion mutants were incubated with recombinant PARP1 protein in the presence of DNA and NAD+. Poly(ADP-ribosyl)ation of GST-FXR mutants was detected by a Western blot assay with an anti-PAR antibody.

Article Snippet: PARP1 activity was assayed using the universal colorimetric PARP assay kit (Trevigen), based on the incorporation of biotinylated ADP-ribose into histone proteins.

Techniques: Immunoprecipitation, Western Blot, Transfection, Recombinant, Incubation, Purification, Staining

Journal: Molecular and Cellular Biology

Article Title: Poly(ADP-Ribose) Polymerase 1 Promotes Oxidative-Stress-Induced Liver Cell Death via Suppressing Farnesoid X Receptor ?

doi: 10.1128/MCB.00160-13

Figure Lengend Snippet: PARP1 bound directly to FXR. (A) Far-Western blot assays of nuclear extracts from HepG2 cells treated with an unrelated siRNA or FXR siRNA. Unpoly(ADP-ribosyl)ated PARP1 (UP-PARP1), autopoly(ADP-ribosyl)ated PARP1 (AP-PARP1), or β-actin protein (negative control) was used as a probe. Histone H1 (his1) served as a loading control (bottom panels). (B) Coimmunoprecipitation assays of FXR-bound proteins from HepG2 cells, followed by Western blot assays using an anti-PARP1 antibody. Nonspecific IgG served as a negative control. (C) Coimmunoprecipitation assays of PARP1-bound proteins or poly(ADP-ribosyl)ated proteins from HepG2 cells, followed by Western blot assays using an anti-FXR antibody. Nonspecific IgG served as a negative control. (D) Far-Western blot assays of recombinant FXR protein. UP-PARP1 or AP-PARP1 was used as a probe. β-Actin protein served as a negative control. (E) Diagram of Flag-tagged human PARP1 with its domains: DNA-binding domain (DBD), nuclear localization signal (NLS), BRCA1 C terminus (BRCT)/automodification domain (AMD), and catalytic domain (CD). Fragments A to F with their amino acid coordinates are listed. HepG2 cells were transfected with EGFP-tagged full-length FXR and Flag-tagged PARP1 mutants. Coimmunoprecipitation assays demonstrated the specific binding of FXR to the BRCT/AMD of PARP1. (F) Diagram of EGFP-tagged human FXR with its domains. LBD, ligand-binding domain. Fragments A to E with their amino acid coordinates are listed. HepG2 cells were transfected with Flag-tagged full-length PARP1 and EGFP-tagged FXR mutants. Coimmunoprecipitation assays demonstrated the specific binding of PARP1 to the LBD of FXR.

Article Snippet: PARP1 activity was assayed using the universal colorimetric PARP assay kit (Trevigen), based on the incorporation of biotinylated ADP-ribose into histone proteins.

Techniques: Far Western Blot, Negative Control, Control, Western Blot, Recombinant, Binding Assay, Transfection, Ligand Binding Assay

Journal: Molecular and Cellular Biology

Article Title: Poly(ADP-Ribose) Polymerase 1 Promotes Oxidative-Stress-Induced Liver Cell Death via Suppressing Farnesoid X Receptor ?

doi: 10.1128/MCB.00160-13

Figure Lengend Snippet: Poly(ADP-ribosyl)ation inhibited the binding of FXR to FXRE in the target promoter. (A) HepG2 cells were treated with 15 μM PJ34 for 24 h with or without H2O2 treatment (300 μM; 12 h). Binding of FXR to FXRE was detected by EMSA. (B) HepG2 cells were transfected with either PARP1 siRNA or an unrelated siRNA at 50 nM for 48 h, with or without H2O2 treatment (300 μM; 12 h). Binding of FXR to FXRE was detected by EMSA. (C) Nuclear extracts from untreated HepG2 cells were incubated with active DNA and NAD+ (1, 10, or 100 μM) and were then subjected to EMSA. (D and E) ChIP-PCR assays using an anti-FXR antibody for amplification of BSEP promoters in HepG2 cells treated with 15 μM PJ34 for 24 h (D) or transfected with PARP1 siRNA or an unrelated siRNA at 50 nM for 48 h (E), with or without H2O2 treatment (300 μM; 12 h). Data are expressed as means ± SEM. Significant differences from the control group (**, P < 0.01) or from the H2O2 group (#, P < 0.05) are indicated. (F) In re-ChIP assays, chromatin was first immunoprecipitated with an anti-FXR or anti-Ctcf (positive-control) antibody and was then reimmunoprecipitated with an anti-PAR antibody, an anti-PARP1 antibody, IgG, or an anti-RNA Pol II antibody. IgG served as a negative control. (G) Nuclear extracts from HepG2 cells were incubated with an anti-FXR, anti-PARP1, or anti-PAR antibody or with nonspecific IgG (negative control) and were then subjected to EMSA. (H) Recombinant FXR proteins were incubated with recombinant PARP1 and/or RXRα as indicated and were then subjected to EMSA.

Article Snippet: PARP1 activity was assayed using the universal colorimetric PARP assay kit (Trevigen), based on the incorporation of biotinylated ADP-ribose into histone proteins.

Techniques: Binding Assay, Transfection, Incubation, Amplification, Control, Immunoprecipitation, Positive Control, Negative Control, Recombinant

Journal: Molecular and Cellular Biology

Article Title: Poly(ADP-Ribose) Polymerase 1 Promotes Oxidative-Stress-Induced Liver Cell Death via Suppressing Farnesoid X Receptor ?

doi: 10.1128/MCB.00160-13

Figure Lengend Snippet: Ligand-induced FXR transactivation was mediated by the inhibition of poly(ADP-ribosyl)ation. (A) Nuclear extracts from HepG2 cells were subjected to an immunoprecipitation assay with an anti-FXR antibody, followed by a Western blot assay using an anti-PAR antibody. Cells were treated with GW4064 (0.5, 1, or 2 μM) or a vehicle (dimethyl sulfoxide) for 24 h. (B) Nuclear extracts from HepG2 cells were subjected to an immunoprecipitation assay with an anti-FXR antibody, followed by a Western blot assay using an anti-PAR antibody. Cells were treated with 1 μM GW4064 for 24 h, with or without H2O2 (300 μM; 12 h). (C) The protein expression of PARP1 or FXR was determined by a Western blot assay. HepG2 cells were treated with GW4064 (0.5, 1, or 2 μM) or a vehicle (dimethyl sulfoxide) for 24 h. (D) EMSAs were used to detect the FXR-FXRE complex in HepG2 cells. After treatment with an empty vector (p3flag-CMV), wt-PARP1, or mut-PARP1 at 1 mg/liter for 24 h, cells were treated with nicotinic acid (50 μM; 24 h), GW4064 (1 μM; 24 h), or CDCA (50 μM; 24 h) as indicated. Nicotinic acid served as a negative control. (E) ChIP-PCR assays using an anti-FXR antibody for the amplification of BSEP promoters. HepG2 cells were treated with an empty vector (p3flag-CMV), wt-PARP1, or mut-PARP1 at 1 mg/liter for 48 h in the absence or presence of nicotinic acid (50 μM; 24 h), GW4064 (1 μM; 24 h), or CDCA (50 μM; 24 h) as indicated. Data are expressed as means ± SEM. Significant differences from the control group (*, P < 0.05) and from the wt-PARP1 transfection group (#, P < 0.05) are indicated.

Article Snippet: PARP1 activity was assayed using the universal colorimetric PARP assay kit (Trevigen), based on the incorporation of biotinylated ADP-ribose into histone proteins.

Techniques: Inhibition, Immunoprecipitation, Western Blot, Expressing, Plasmid Preparation, Negative Control, Amplification, Control, Transfection

Journal: PeerJ

Article Title: Effects of poly (ADP-ribose) polymerase-1 (PARP-1) inhibition on sulfur mustard-induced cutaneous injuries in vitro and in vivo

doi: 10.7717/peerj.1890

Figure Lengend Snippet: Cultured HaCaT cells were treated with various concentrations of SM for 6 h. The level of pADPr protein was determined by immunofluorescence (A and B), western blotting (C and D) and Acumen (E), respectively. (A and B) The scale shown in the upper left panel (bar = 20 µm) was the same for all panels. (C and D) The results of the western blots were normalized to the levels of GAPDH and then presented as the fold of the control levels. (E) The results of the Acumen analysis were normalized to the nuclear total fluorescence intensity. ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. untreated group. ## P < 0.01, ### P < 0.001 vs. SM-treated group. (F) The PARP-1 inhibitory effect of ABT-888 was also confirmed at enzyme level. (Each data point represents three data points).

Article Snippet: The PARP-1 inhibitory effect of ABT-888 was also confirmed using the Trevigen’s Homogeneous PARP Inhibition Assay Kit ( ).

Techniques: Cell Culture, Immunofluorescence, Western Blot, Control, Fluorescence

Journal: PeerJ

Article Title: Effects of poly (ADP-ribose) polymerase-1 (PARP-1) inhibition on sulfur mustard-induced cutaneous injuries in vitro and in vivo

doi: 10.7717/peerj.1890

Figure Lengend Snippet: (A) Hematoxylin and eosin stains of the mouse ear skin showed that the PARP inhibitor ABT-888 did not have a protective effect against pathological damage in mice exposed to 0.16 mg SM/ear, but ABT-888 produced reductions in pathological damage in mice exposed to 0.64 mg SM/ear. (A1) The medial surface of a normal ear from the control group. (A2) The medial surface of an ear from the 0.16 mg SM/ear exposure group, showing epidermal necrosis (pyknotic nuclei, arrowhead). (A3) The medial surface of an ear from the 0.16mg SM/ear exposure + ABT-888 group. ABT-888 showed no protective effect. (A4) The medial surface of an ear from the 0.64 mg SM/ear exposure group. Epidermal necrosis (arrowhead) was progressively more severe. (A5) The medial surface of an ear from the 0.64 mg SM/ear exposure + ABT-888 group. ABT-888 could reduce reticular degenerative changes in the dermis, hypereosinophilic cytoplasms of the epidermis necrosis (arrowhead). The scale shown in the lower right panel (bar = 20 µm) is the same for all panels. (B and C) ABT-888 significantly reduced edema (REW, approximately 26%) of the ear and epidermal necrosis (EN score, approximately 40%) in MEVM in the group exposed to 0.64 mg SM/ear, but showed no protective effect in the group exposed to 0.16 mg SM/ear. ABT-888 was administered (i.p.) 30 min before the SM exposure. ∗ P < 0.05 vs. 0.64 mg SM/ear. All the data are presented as means ±SEM ( n = 5–7).

Article Snippet: The PARP-1 inhibitory effect of ABT-888 was also confirmed using the Trevigen’s Homogeneous PARP Inhibition Assay Kit ( ).

Techniques: Produced, Control

Journal: PeerJ

Article Title: Effects of poly (ADP-ribose) polymerase-1 (PARP-1) inhibition on sulfur mustard-induced cutaneous injuries in vitro and in vivo

doi: 10.7717/peerj.1890

Figure Lengend Snippet: Caspase 3/7 activity was measured 6 h (A) and 24 h (B) after SM exposure and the treatment of ABT-888. Total cell lysates were prepared and analyzed for protein expression by western blotting using anti-PARP-1 and cleavage of PARP-1 antibody 6 h (C) and 24 h (D) after SM exposure and the treatment of ABT-888. β -Actin was used as a control to ensure equal protein loading. Apoptosis and necrosis were analyzed by flow cytometry 6 h (E) and 24 h (F) after SM exposure and the treatment of ABT-888. The frames were divided into four quadrants: Annexin V−/PI− normal cells are in quadrant I; Annexin V+/PI− apoptotic cells are in quadrant II; PI+ necrotic cells are in quadrant III/IV. The values are presented as means ± SEM, n = 3 or 6. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. untreated group. ## P < 0.01, ### P < 0.001 vs. SM-treated group.

Article Snippet: The PARP-1 inhibitory effect of ABT-888 was also confirmed using the Trevigen’s Homogeneous PARP Inhibition Assay Kit ( ).

Techniques: Activity Assay, Expressing, Western Blot, Control, Flow Cytometry

Journal: PeerJ

Article Title: Effects of poly (ADP-ribose) polymerase-1 (PARP-1) inhibition on sulfur mustard-induced cutaneous injuries in vitro and in vivo

doi: 10.7717/peerj.1890

Figure Lengend Snippet: To evaluate whether PARP-1 was involved in the toxicity of SM, Lv-shPARP-1 and Lv-shCon were transfected into HaCaT cells; then, stable transfectants were selected. The efficacy for PARP-1 knockdown was determined by RT-PCR (A) and western blotting (B). The control and PARP-1 knockdown HaCaT cells were treated with 0, 100, or 1,000 µM SM. Subsequently, the cell viability was measured 6 h (C) and 24 h (D) after exposure to SM. The results are presented as means ± SEM determined from three independent experiments. ∗∗∗ P < 0.001 vs. untreated group. # P < 0.05, ### P < 0.001 vs. SM-treated group.

Article Snippet: The PARP-1 inhibitory effect of ABT-888 was also confirmed using the Trevigen’s Homogeneous PARP Inhibition Assay Kit ( ).

Techniques: Transfection, Knockdown, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: PeerJ

Article Title: Effects of poly (ADP-ribose) polymerase-1 (PARP-1) inhibition on sulfur mustard-induced cutaneous injuries in vitro and in vivo

doi: 10.7717/peerj.1890

Figure Lengend Snippet: PARP-1-knockdown and control HaCaT cells were treated with 100 µM or 1,000 µM SM. At 6 h and 24 h after exposure to SM, the cells were harvested for the detection of the apoptosis checkpoint signals. The protein levels of phospho-JNK (Thr183/Tyr185) 6 h (A) and 24 h (B) after exposure to SM, phospho-p53 (ser46) 6 h (C) and 24 h (D) after exposure to SM, active Caspase 9 (Asp315) 6 h (E) and 24 h (F) after exposure to SM, active Caspase 8 (Asp384) 6 h (G) and 24 h (H) after exposure to SM, and c-PARP (p89) 6 h (K) and 24 h (L) after exposure to SM were determined using Luminex assays. The caspase 3/7 activity 6 h (I) and 24 h (J) after exposure to SM was measured using the Caspase-Glo 3/7 assay kit. The results are presented as means ±SEM, as determined from three independent experiments. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. untreated group. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. SM-treated group.

Article Snippet: The PARP-1 inhibitory effect of ABT-888 was also confirmed using the Trevigen’s Homogeneous PARP Inhibition Assay Kit ( ).

Techniques: Knockdown, Control, Luminex, Activity Assay, Caspase-Glo Assay

Journal: PeerJ

Article Title: Effects of poly (ADP-ribose) polymerase-1 (PARP-1) inhibition on sulfur mustard-induced cutaneous injuries in vitro and in vivo

doi: 10.7717/peerj.1890

Figure Lengend Snippet: PARP-1-knockdown and control HaCaT cells were treated with 100 µM or 1,000 µM SM. At 6 h and 24 h after exposure to SM, the cells were harvested for the detection of Phospho-AKT (Thr308) (A, B) and Phospho-mTOR (Ser2448) (C, D). The results are presented as means ± SEM, as determined from three independent experiments. ∗ P < 0.05, ∗∗ P < 0.01, vs. untreated group. # P < 0.05, ## P < 0.01, vs. SM-treated group.

Article Snippet: The PARP-1 inhibitory effect of ABT-888 was also confirmed using the Trevigen’s Homogeneous PARP Inhibition Assay Kit ( ).

Techniques: Knockdown, Control

Journal: PeerJ

Article Title: Effects of poly (ADP-ribose) polymerase-1 (PARP-1) inhibition on sulfur mustard-induced cutaneous injuries in vitro and in vivo

doi: 10.7717/peerj.1890

Figure Lengend Snippet: Schematic of the role of PARP-1 in sulfur mustard injury.

Article Snippet: The PARP-1 inhibitory effect of ABT-888 was also confirmed using the Trevigen’s Homogeneous PARP Inhibition Assay Kit ( ).

Techniques:

Journal: Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer

Article Title: Positron-Emission Tomographic Imaging of a Fluorine 18-Radiolabeled Poly(ADP-Ribose) Polymerase 1 Inhibitor Monitors the Therapeutic Efficacy of Talazoparib in SCLC Patient-Derived Xenografts.

doi: 10.1016/j.jtho.2019.05.032

Figure Lengend Snippet: Figure 2. Fluorine 18 poly(ADP-ribose) polymerase inhibitor ([18F]PARPi) positron-emission (PET)/computerized tomography (CT) experimental design. (A) Molecular structures of olaparib, talazoparib, and an [18F]PARPi are shown. The [18F]PARPi is formed by conjugating a 4-[18F]fluorobenzoic acid group to a scaffold of the small molecule olaparib. (B) When [18F]PARPi is present without the competitive binding of talazoparib, it binds to the PARP1 enzyme, allowing visualization of the high PARP expressing SCLC patient-derived xenograft SCRX-Lu149. Tumor is marked with a “T” on PET images. Tumors are implanted in the right shoulder to prevent signal interference from the gut, as the [18F]PARPi radiotracer is eliminated by the hepatobiliary system and subsequently has increased activity in the intestines. (C) Pre-treatment with 0.2 mg/kg talazoparib by oral gavage competitively blocks the binding of [18F]PARPi and results in lower activity. (D) An experimental timeline shows the method used for talazoparib pre-treatment and PET scanning. Talazoparib was orally dosed at 0.1, 0.2, or 0.3 mg/kg at various timepoints before intravenous injection of [18F]PARPi. Two hours after radiotracer injection, PET/CT imaging was performed, and mouse dissection was performed immediately after imaging. IV, intravenous; ID, injected dose; PAR, poly(ADP-ribose); ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Tumors were lysed and poly-(ADP) ribose (PAR) values were quantified using the PARP in vivo Pharmacodynamic Assay II Kit (Trevigen, Gaithersburg, Maryland).

Techniques: Tomography, Binding Assay, Expressing, Derivative Assay, Activity Assay, Injection, Positron Emission Tomography-Computed Tomography, Imaging, Dissection, Enzyme-linked Immunosorbent Assay