β tubulin t4026

Proteins were extracted with RIPA buffer (RIPA buffer, 1 mM DTT, 1M NaF, 100 mM sodium orthovanadate and protease and phosphatase inhibitors). After SDS PAGE separation, and transfer onto a PVDF membrane by iBlot dry blotting system (Invitrogen), membranes were incubated overnight at 4°C with primary antibodies: HER2 (GTX50425; Genetex), βIII-tubulin (clone TUJ1), βII-tubulin (clone 7B9) α-tubulin (T6199), β-tubulin (T4026) and β-actin (A5441) from Sigma-Aldrich, and 1h at room temperature with secondary antibodies (IRDye Infrared Dyes from LI-COR Biosciences). Membranes were scanned using Odyssey infrared imaging system (LI-COR Biosciences) and densitometric quantification was performed with Odyssey software. Quantification of Western Blot was performed using Image J software (Rasband, W.S., National. Institutes of Health). Expression levels of proteins were normalized against β-actin.

N-acetyl-L-cysteine(NAC), hydrogen peroxide(H₂O₂), MTT [3(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and 4′,6-diamidno-2-phenylinole(DAPI) were purchased from the Sigma Chemical Co. (St. Louis, MO, USA). 2’,7’–dichlorofluorescin diacetate (DCF-DA) was purchased from Molecular Probe (Eugene, Oregon, USA). Rabbit antibodies which are reactive with Nrf2 (12,721) were from Cell Signaling Technology Inc. (Danvers, MA, USA). Mouse antibodies which are reactive with acetylated tubulin (T7451) or β-tubulin (T4026) were from Sigma-Aldrich Co. (St. Louis, MO, USA). Rabbit antibodies which are reactive with Arl13b (17,711–1-AP) were from Proteintech Group Inc. (Rosemont, IL, USA). Chicken anti-mouse IgG-Alexa 488 (A-21200) and goat anti-rabbit-Alexa 568 (A-11011) were obtained from Invitrogen (Calsbad, CA, USA). Except where indicated, all other materials are obtained from the Sigma Chemical Co. (St. Louis, MO, USA)^{17 (link)}.

Dimethyloxallyl Glycine (DMOG, 71210) was from Cayman Chemical (Ann Arbor, MI, USA). N-acetyl-L-cysteine (NAC), hydrogen peroxide (H₂O₂), MTT [3(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide], sodium butyrate and 4′,6-diamidno-2-phenylinole (DAPI) were purchased from the Sigma Chemical Co. (St. Louis, MO, USA). 2’,7’-dichlorofluorescin diacetate (DCF-DA) was purchased from Molecular Probe (Eugene, OR, USA). Ciliobrevin A was from Selleck Chemicals (Houston, TX, USA). PD98059 was from Enzo Life Sciences, Inc. (Farmingdale, NY, USA). Mouse antibodies which are reactive with acetylated tubulin (T7451), and β-tubulin (T4026) were from Sigma-Aldrich Co. (St. Louis, MO, USA). Rabbit antibodies which are reactive with Nrf2 and HIF-1α were from Cell signaling Technology (Cat# 12721, Danvers, MA, USA) and Novus Biologicals (Centennial, CO, USA), respectively. Rabbit antibodies which are reactive with Arl13b (17711-1-AP) were from Proteintech Group Inc. (Rosemont, IL, USA). Chicken anti-mouse IgG-Alexa 488 (A-21200) and goat anti-rabbit-Alexa 568 (A-11011) were obtained from Invitrogen (Calsbad, CA, USA). Except where indicated, all other materials are obtained from the Sigma Chemical Co. (St. Louis, MO, USA).