parp inhibitors 3-ab  (Porsolt)

Journal: Frontiers in Oncology

Article Title: Actinomycin D-Activated RNase L Promotes H2A.X/H2B-Mediated DNA Damage and Apoptosis in Lung Cancer Cells

doi: 10.3389/fonc.2019.01086

Figure Lengend Snippet: Actinomycin D (Act D) promotes RNase L dimerization and RNase L-dependent apoptosis in lung cancer cells. (A,B) Western blot of dose dependence (10 −3 -10 2 μg/ml) (0.8–8 × 10 4 nM) (A) and time course (B) of Act D-induced RNase L dimerization. (C) Cleavage activity of RNase L and Western blot of activated Caspase-3 and PARP after Act D (0.1 μg/ml) (80 nM) treatment in the indicated time. (D) Cleavage activity of RNase L, Western blot of RNase L dimerization and activated Caspase-3 and PARP after RNase L-knock down (si-RNase L) or Caspase-3 inhibitor (z-DEVD-fmk) with the treatment of Act D (0.1 μg/ml) (80 nM) for 24 h. Data were collected from triplicate samples in three independent experiments. ★ P < 0.05; **, ♦♦, ★★ P < 0.01;***, ♦♦♦, ★★★ P < 0.001.

Article Snippet: Caspase-3 inhibitor (z-DEVD-fmk), PARP inhibitor (3-AB), and ROCK-1 inhibitor (Y-27632) were purchased from Calbiochem, USA.

Techniques: Western Blot, Activity Assay, Knockdown

Journal: Frontiers in Oncology

Article Title: Actinomycin D-Activated RNase L Promotes H2A.X/H2B-Mediated DNA Damage and Apoptosis in Lung Cancer Cells

doi: 10.3389/fonc.2019.01086

Figure Lengend Snippet: RNase L induces DNA cleavage and condensation with H2A.X-H2B activation through Caspase-3/PARP activation. (A) Immunocytofluorescence (ICF) of p-Ser139-H2A.X, TUNEL, and DAPI in NCI-H460 treated with Act D and Caspase-3 inhibitor (z-DEVD-fmk) or PARP inhibitor (3-AB). (B) Western blot of p-Ser139-H2A.X and H2A.X after RNase L interference. (C) ICF of DAPI in NCI-H460 treated with Act D in the indicated time. (D) Western blot of RNase L, p-Ser14-H2B, and H2B treated with Act D in the indicated time after RNase L interference. (E) Western blot of p-Ser14-H2B and H2B treated with Act D and Caspase-3 inhibitor (z-DEVD-fmk) or PARP inhibitor (3-AB). Data were collected from triplicate samples in three independent experiments. *** P < 0.001.

Article Snippet: Caspase-3 inhibitor (z-DEVD-fmk), PARP inhibitor (3-AB), and ROCK-1 inhibitor (Y-27632) were purchased from Calbiochem, USA.

Techniques: Activation Assay, TUNEL Assay, Western Blot

Journal: Frontiers in Oncology

Article Title: Actinomycin D-Activated RNase L Promotes H2A.X/H2B-Mediated DNA Damage and Apoptosis in Lung Cancer Cells

doi: 10.3389/fonc.2019.01086

Figure Lengend Snippet: RNase L induces apoptosis through Caspase-3/ROCK-1/PARP/H2A.X-H2B/p21 cascade. (A–C) ICF of Annexin V and PI (DNA) (A) and analysis of Annexin V + (B) or PI + (C) in NCI-H460 in the presence of Act D with or without ROCK-1 inhibitor (Y-27632) for the indicated time. (D) Western blot of RNase L and ROCK-1 in NCI-H460 treated with Act D after RNase L interference. (E) Western blot of Caspase-3, ROCK-1, and PARP in the presence of Act D with or without Caspase-3 inhibitor (z-DEVD-fmk) or ROCK-1 inhibitor (Y-27632). (F) Western blot of p21 in NCI-H460 treated with Act D and different intervention [siRNA interference for RNase L, H2A.X, H2B or H2A.X, and H2B combined, Caspase-3 inhibitor (z-DEVD-fmk), ROCK-1 inhibitor (Y-27632), and PARP inhibitor (3-AB)]. (G) Graphical summary for Act D-activated RNase L-mediated apoptosis through Caspase 3-ROCK 1-PARP-H2A.X/H2B-p21 cascade. Data were collected from triplicate samples in three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Caspase-3 inhibitor (z-DEVD-fmk), PARP inhibitor (3-AB), and ROCK-1 inhibitor (Y-27632) were purchased from Calbiochem, USA.

Techniques: Western Blot

Journal: International Journal of Neuropsychopharmacology

Article Title: Antidepressant-Like Actions of Inhibitors of Poly(ADP-Ribose) Polymerase in Rodent Models

doi: 10.1093/ijnp/pyx068

Figure Lengend Snippet: Effect of poly(ADP-ribose) polymerase (PARP) inhibitors on immobility time (A) and latency to immobility (B) in the Porsolt swim test. Rats were treated daily for 10 days prior to the swim test with either saline (vehicle i.p.: n=13), 3-aminobenzamide (3-AB) administered at 3 different doses as noted (s.c.; n=8-10), or 5-AIQ (0.3 mg/kg i.p.; n=9). Additional groups of rats were administered 3 injections over 24 hours prior to the swim test with either fluoxetine (10 mg/kg i.p. per injection; n=10; FLX x 3) or 3-AB (40 mg/kg s.c. per injection; n=7; 3-AB x 3). The swim test data were collected 2 h after the final drug or vehicle injection. Asterisks indicate significant differences compared to the vehicle group (* P <.05, ** P <.01). The results of statistical analyses of all other comparisons can be found in Supplemental Table 1.

Article Snippet: Two structurally different PARP inhibitors, 3-AB and 5-AIQ, demonstrated antidepressant-like activity in the Porsolt swim test.

Techniques: Saline, Injection

Journal: Scientific Reports

Article Title: miR-7a/b attenuates post-myocardial infarction remodeling and protects H9c2 cardiomyoblast against hypoxia-induced apoptosis involving Sp1 and PARP-1

doi: 10.1038/srep29082

Figure Lengend Snippet: ( A ) Immunostaining of PARP-1 in vivo . ( B ) Western blots of PARP-1 in vivo . ( C ) Western blots of PARP-1 in cells exposed to hypoxia for different time. ( D–G ) Western blots showing the effect of miR-7a/b on regulation of PARP-1 ( D , E ), cleaved caspase-3 ( D,F ), Bax/Bcl-2 ( D , G ). ( H,I ) TUNEL assay results (Scale bar: 50 μm). Con: sham mice without LAD occlusion ( A,B ) or normal cultured H9c2 cells ( C–H ). MI: mice with LAD occlusion. NC: H9c2 cells exposed to hypoxia for 12 h. Data are the mean ± SD, n = 6/group ( A ), n = 3/group ( B–I ), *p < 0.05 compared with Con; # p < 0.05 compared with GFP-NC ( A,B ) or NC ( C–I ).

Article Snippet: The cells were treated with or without PARP-1 inhibitor 3-AB (3 mM) (Sigma) or the Sp1 inhibitor mithramycin (Cayman Chemical) before being placed in the Whitley H35 Hypoxystation (Don Whitley Scientific) for hypoxia (1% O 2 , 5% CO 2 ) treatment.

Techniques: Immunostaining, In Vivo, Western Blot, TUNEL Assay, Cell Culture

Journal: Scientific Reports

Article Title: miR-7a/b attenuates post-myocardial infarction remodeling and protects H9c2 cardiomyoblast against hypoxia-induced apoptosis involving Sp1 and PARP-1

doi: 10.1038/srep29082

Figure Lengend Snippet: ( A–D ) Western blots showing the effect of 3-AB on regulation of PARP-1 ( A,B ), cleaved caspase-3 ( A,C ), Bax/Bcl-2 ( A,D ). ( E,F ) TUNEL assay results (Scale bar: 50 μm). Con: normal cultured H9c2 cells. –, cells only exposed to hypoxia. Data are the mean ± SD, n = 3/group, *p < 0.05 compared with Con; # p < 0.05 compared with-.

Article Snippet: The cells were treated with or without PARP-1 inhibitor 3-AB (3 mM) (Sigma) or the Sp1 inhibitor mithramycin (Cayman Chemical) before being placed in the Whitley H35 Hypoxystation (Don Whitley Scientific) for hypoxia (1% O 2 , 5% CO 2 ) treatment.

Techniques: Western Blot, TUNEL Assay, Cell Culture

Journal: Scientific Reports

Article Title: miR-7a/b attenuates post-myocardial infarction remodeling and protects H9c2 cardiomyoblast against hypoxia-induced apoptosis involving Sp1 and PARP-1

doi: 10.1038/srep29082

Figure Lengend Snippet: ( A,D–F ) Western blots of Sp1, PARP-1 and caspase-3 in hypoxic cells pretreated with different concentration of mithramycin (nM). ( C,G–I ) Western blots of Sp1, PARP-1 and caspase-3 in hypoxic H9c2 cells transfected with miR-7a/b inhibitors that treated with or without 100 nM mithramycin. ( B ) Representative ChIP assays. Con: normal cultured H9c2 cells, M: mithramycin, *p < 0.05 compared with control group, # p < 0.05 compared with NC siRNA-transfected group, ♦ p < 0.05 compared with miR-7a/b inhibitors-transfected group; Φ p < 0.05 compared with mithramycin-treated group.

Article Snippet: The cells were treated with or without PARP-1 inhibitor 3-AB (3 mM) (Sigma) or the Sp1 inhibitor mithramycin (Cayman Chemical) before being placed in the Whitley H35 Hypoxystation (Don Whitley Scientific) for hypoxia (1% O 2 , 5% CO 2 ) treatment.

Techniques: Western Blot, Concentration Assay, Transfection, Cell Culture