Anti nurr1

Angiotensin II was purchased from Sigma-Aldrich (St Louis, MO, USA). AG14361 was purchased from Selleck (Houston, TX, USA) and dissolved in Dimethyl sulfoxide (DMSO). siRNAs were purchased from GE Healthcare (Uppsala, Sweden). Following antibodies were used: anti-Nurr1/Nur77 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA, sc-990) and anti-Nurr1 (Santa Cruz Biotechnology, Inc., sc-991) for RIME, anti-Nurr1 (Perseus Proteomics, Tokyo, Japan, PP-N1404-00), anti-PARP1 (Transduction Laboratories, Lexington, KY, USA, P76420), anti-FLAG tag (Sigma-Aldrich, F1804) and anti-HA tag (ICL, Portland, OR, USA, RHGT-45A-Z) for western blot, anti-Nurr1 (Santa Cruz Biotechnology, Inc., sc-991) for immunoprecipitation, anti-Nurr1 (Atlas Antibodies, Bromma, Sweden, HPA000543) and anti-PARP1 (Transduction Laboratories, P76420) for immunostaining, Alexa Fluor 488 goat anti-rabbit IgG (H + L) (Thermo Fisher Scientific, Pittsburgh, PA, USA, A11034) and Alexa Fluor 594 goat anti-mouse IgG (H + L) (Thermo Fisher Scientific, A11032) for 2nd antibodies for immunostaining. For immunoprecipitation of FLAG-tagged protein, Anti-FLAG M2 affinity agarose gel (Sigma-Aldrich, A2220) were used.

H295R cells were fixed with 3.5% formaldehyde for 15 min at room temperature. After blocking with 1% skim milk, cells were incubated with indicated primary antibodies for 1 h at room temperature. The cells were subsequently washed and incubated with Alexa-coupled 2nd antibodies. Images of stained cells were taken using an LSM800 confocal microscope (Zeiss, Oberkochen, Germany) Dilution rates of antibodies used in this study were as follows: anti-Nurr1 (Atlas Antibodies) and anti-PARP1 (Transduction Laboratories), 1/100, and Alexa Fluor 488 goat anti-rabbit IgG (H + L) and Alexa Fluor 594 goat anti-mouse IgG (H + L), 1/500.