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: