Poly adp ribose polymerase parp

Antibody targeting TXNRD1 (cat. no. ab16840) was purchased from Abcam (Cambridge, UK), and poly (ADP-ribose) polymerase (PARP; cat. no. 9532s), cleaved-PARP (cat. no. 9542) and apoptosis regulator Bax (cat. no. 5023), were from Cell Signaling Technology, Inc. (Danvers, MA, USA). Anti- nuclear factor erythroid 2-related factor 2 (NRF2) antibody (cat. no. sc-81342) was purchased from Santa Cruz Biotechnology (Dallas, TX. USA). Anti-β-actin antibody (cat. no. MABT825) was purchased from Sigma-Aldrich (Merck Millipore). Fluorescence-labeled secondary antibody (IRDye^® 800CW; cat. no. 925-32211) was purchased from LI-COR Biosciences (Lincoln, NE, USA). Protein lysates were prepared with radioimmunoprecipitation assay buffer and quantified using the Bradford method with a Bradford Protein Assay kit purchased from Bio-Rad Laboratories, Inc. A total of ~40 µg total protein was used for western blot analysis. Proteins were separated using SDS-PAGE on a 10% gel and subsequently transferred to a polyvinylidene fluoride membrane. Membranes were incubated with primary antibodies at 4°C overnight and with secondary antibodies at room temperature for 2 h. Protein band images were captured using an Odyssey Imaging scanner from LI-COR Biosciences.

Collected MPG and PN, and bladder were fixed at 4 °C in 4% paraformaldehyde for 24 hours to make paraffin blocks. The primary antibodies used were as follows: βIII-tubulin (diluted 1:200; Abcam, Cambridge, MA, USA), brain-derived neurotrophic factor (BDNF diluted 1:100; Abcam, Cambridge, MA, USA), poly-ADP-ribose polymerase (PARP, diluted 1:500; Abcam, Cambridge, MA, USA) were used. 6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Inc., Burlingame, CA, USA) was used to stain nuclei. The digital images were obtained by using Zeiss LSM 510 Meta confocal microscope (Zeiss, Oberkochen, Germany). Mean intensity was calculated using ZEN 2012 (Zeiss).

The collected PN and bladder were fixed in 4% paraformaldehyde for 24 h at 4°C
before creating a paraffin block, as described previously⁹ . The primary antibodies were used as follows: vascular endothelial growth
factor (VEGF; diluted 1:200; Santa Cruz Biotechnologies, Santa Cruz, CA, USA),
βIII-tubulin (diluted 1:200; Abcam, Cambridge, UK), bFGF (diluted 1:500; Cell
Signaling Technology, Danvers, MA, USA), SDF-1 (diluted 1:200; Abcam, Cambridge,
UK), poly-ADP-ribose polymerase (PARP, diluted 1:500; Abcam, Cambridge, UK), and
6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Inc., Burlingame, CA,
USA) were used to stain the nuclei. Digital images were obtained using a Zeiss
LSM 510 Meta confocal microscope (Zeiss, Oberkochen, Germany), and bladder
images were analyzed using ZEN 2012 (Zeiss, Oberkochen, Germany). The other
resulting images were analyzed using Image J to determine the positive rate for
each figure. In brief, we used Image J to split each color from merged figure,
calculate the intensity of each color, and then added them. The positive was the
ratio of a special color fluorescence intensity to all colors intensity.

Propofol (Fresenius Kabi Austria Gmbh, Hafnerstrasse, Austria) diluted with dimethyl sulfoxide (DMSO), Hoechst 33342, and chloroquine were purchased from Sigma (St. Louis, MO, USA). The following reagents were obtained commercially: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), acridine orange (AO), monodansylcadaverine (MDC), and 3-methyladenine (3-MA, class III PI3K inhibitor) (Calbiochem, La Jolla, CA, USA). The antibodies used in the study were as follows: Bak (1:1000), caspase-9 (1:1000), caspase-3 (1:1000), and poly (ADP-ribose) polymerase (PARP; 1:1,000) (Abcam, Cambridge, MA, USA).