Phospho retinoblastoma protein rb

Amlodipine and gefitinib were purchased from Selleck Chemicals (Houston, TX, United States) and TopScience (Shanghai, China), respectively. Propidium iodide (PI) was obtained from Sigma-Aldrich (St. Louis, MO, United States). The Annexin V/FITC-PI apoptosis detection kit was obtained from BD Biosciences (San Jose, CA, United States of America). RPMI 1640 and FBS were purchased from Biological Industries (Beit Haemek, Israel). The enhanced chemiluminescence (ECL) reagent was purchased from Thermo Fisher Scientific (Waltham, MA, United States). Antibodies against phospho-EGFR, phospho-PDK1 (Ser241), PDK1, phospho-Akt (Ser473), phospho-Akt (Thr308), Akt, phospho-mammalian target of rapamycin complex 1 (mTORC1) (Ser2448), mTOR, phospho-glycogen synthase kinase 3 (GSK-3) β, phospho-p70 S6K (Thr389), p27, cyclin D1, phospho-retinoblastoma protein (Rb), phospho-extracellular signal-regulated kinase (ERK) 1/2, phospho-c-Raf, β-actin, and horseradish peroxidase (HRP)-conjugated goat anti-rabbit and horse anti-mouse secondary antibodies were purchased from Cell Signaling Technology Inc. (Danvers, MA, United States). Antibody specific for p21 was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, United States).

Chromatographically pure Sch was purchased from Feng-Shan-Jian Medicine Research Co. Ltd. (Kunming, Yunnan, China) (Structure shown in Figure 1), and dissolved in 0.5% dimethyl sulfoxide (DMSO) at a concentration of 0.5 mg/mL before experiment. LPS (Escherichia coli serotype O55:B5), Evans blue (EB) dye, and rhodamine 6G were purchased from Sigma Chemical (St. Louis, MO, United States). Primary antibodies against I-κBα, phospho-I-κBα, NF-κB p65, Erk1/2, phospho-Erk1/2, p38 MAPK, phospho-p38 MAPK, phospho-FoxO1, Akt, phospho-Akt, Keratin, caspase-3, cyclin D1, cyclin B1, p27^kip1, phospho-retinoblastoma protein (Rb), GAPDH, and histone H3 were obtained from Cell Signaling Technology (Beverly, MA, United States). Antibodies against Claudin-5, Occludin and ZO-1 were from Invitrogen (Rochford, IL, United States), antibodies against FoxO1, p21^cip1, myeloperoxidase (MPO) and von Willebrand factor (vWF) from Abcam (Cambridge, United Kingdom). Antibody against VE-Cadherin from Santa Cruz Biotechnology (Santa Cruz, CA, United States), antibody against TLR4 from Novus Biologicals (Littleton, CO, United States) and antibody against prosurfactant protein C (proSP-C) from EMD Millipore Corporation (Temecula, CA, United States).