Nqo 1 antibody

Lung tissue and cells were plunged in ice-cold RIPA lysis buffer with protease and phosphatase inhibitors (Abcam, Cambridge, UK). Per the manufacturer's instructions, protein concentrations were quantified using the BCA Protein Assay Kit (Beyotime Biotech, Shanghai, China). Briefly, protein samples were loaded and separated on SDS-PAGE gels, transferred to PVDF membranes (Bio-Rad, USA), and blocked with 5% (w/v) skimmed milk containing 0.1% PBS buffer Tween 20 (v/v) or 5% (w/v) protease free bovine serum Albumin (BSA) (Sigma-Aldrich, USA). Then the membranes were incubated with β-actin antibody (1:5000, Cat#: 81,115-1-RR, SAB), NLRP3 anti-body (1:2000, Cat#: 15,101, CST), pro-caspase-1/p10/p20 antibody (1:1000, Cat#: ab179515, Abcam),Heme Oxygenase 1 antibody (1:1000, Cat#: ab52947, Abcam), NQO1 antibody (1:3000, Cat#: ab80588, Abcam),p-GSK3β (S9) antibody (1:1000, Cat#: WL03518, WanleiBio), GSK3β antibody (1:300, Cat#: WL01456, WanleiBio), p-AKT (Ser473) antibody (1:500, Cat#: WLP001a, WanleiBio), AKT antibody (1:500, Cat#: WL0003b, WanleiBio)overnight at 4 ℃, and secondary antibodies (1: 5000, SAB) were labeled with horseradish peroxidase for 2 h at room temperature. After washing with PBST, bands were detected with Luminata^™ Crescendo chemiluminescence horseradish peroxidase substrate (Millipore, USA) and scanned using GeneGnome XRQ imager (Syngene, UK).

MPS was purchased from Pfizer and dissolved in sterile normal saline. VO-OHpic was purchased from selleck (USA, S8174) and dissolved in DMSO. And the final concentration of DMSO is the same (0.1%) in each group including the control. CD31 antibody (UK, ab182981), CD34 antibody (UK, ab81289), VEGF Receptor 2 (VEGFR2) antibody (UK, ab11939), VEGF Receptor 1 (VEGFR1) antibody (UK, ab32152), CD133 antibody (UK, ab19898), SIRT-1 antibody (UK, ab189494), NQO-1 antibody (UK, ab80588), and Trx antibody (UK, ab133524) were purchased from Abcam. FITC-conjugated mouse anti-rabbit IgG antibody was purchased from Boster (China, BM2012). Bad antibody (USA, #9268, for IP), Phospho-Bad antibody (USA, #5284), Bcl-xL antibody (USA, #2764), AKT antibody (USA, #4691), p-AKT antibody (USA, #4060), cleaved caspase-3 (Asp175) antibody (USA, #9661), cleaved caspase-9 antibody (USA, #9508), MFF antibody (USA, #84580), and normal rabbit IgG (USA, #2729) were purchased from Cell Signaling Technology. Bax antibody (USA, 50599-2-Ig, for western blot), Cytochrome C antibody (USA, 66264-1-Ig), Nrf2 antibody (USA, 16396-1-AP), Bcl-xL antibody (USA, 26967-1-AP), Bad antibody (USA, 10435-1-AP), FIS1 antibody (USA, 10956-1-AP), VEGF antibody (USA, 19003-1-AP), HO-1 antibody (USA, 10701-1-AP), and GAPDH antibody (USA, 60004-1-Ig) were purchased from Proteintech.

The human pancreatic cancer cell lines PANC-1 and CFPAC-1 used in our study were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and stably cultured, passaged, and cryopreserved in our laboratory. Basic Dulbecco's modified Eagle's medium (DMEM, Gibco, USA) with 10% fetal bovine serum (FBS, Sigma, St. Louis, MO, USA) was utilized to culture PANC-1 cells, while basic RPMI-1640 medium (Gibco, USA) with 10% FBS (Sigma, St. Louis, MO, USA) was utilized to culture CFPAC-1 cells, which were cultured in an incubator at 37°C, 5% CO₂, and 95% humidity. The drug curcumin was purchased from MedChemExpress (MCE, New Jersey, USA). The anti-sestrin2 antibody was purchased from Proteintech (Chicago, USA), while the anti-Nrf2 antibody, p-Nrf2 antibody, Keap1 antibody, HO-1 antibody, and NQO-1 antibody were purchased from Abcam (Britain).