The Roswell Park Memorial Institute (PRMI 1640) medium was purchased from Gibco (Life Technologies

Corporation, Grand Island, NY, USA). Sodium dihydrogen phosphate, sodium chloride, sucrose, and other chemicals were purchased from the Chinese histone deacetylase activity Medicine Group Chemical Reagent Corporation (Shanghai, China). Micro bicinchoninic acid (Micro BCA) protein kit was purchased from Pierce Biotechnology, Inc. (Vallejo, CA, USA). Preparation of dextran nanoparticles loaded with C188-9 cost proteins The model proteins, BSA, GM-CSF, β-galactosidase, and MYO were encapsulated into dextran nanoparticles according to aqueous-aqueous freezing-induced phase separation methods. Briefly, proteins were dissolved in 6% (w/w) dextran solutions as separated phase, and the polyethylene glycol (PEG) was dissolved to get an aqueous solution with a concentration of 6% (w/w). Then, the two solutions were gently mixed to get a clear solution. The solution was frozen at −80°C in the refrigerator for more than 10 h and then dried at a vacuum level below 0.1 mbar for 24 h. After lyophilization,

the powder was washed PARP inhibitor with dichloromethane and subsequently centrifuged at 12,000 rpm for 3 min and three times to remove the continuous phase. Once dichloromethane was evaporated, fine dextran nanoparticles loaded with proteins were obtained. Morphology of dextran nanoparticles loaded with proteins The morphology analysis was measured by scanning electron

microscopy (SEM). The dextran nanoparticles were attached to a metal stub using a double-sided adhesive and exposed to gold spray under argon atmosphere for 10 min. The size distribution of dextran nanoparticles was measured using a photon correlation spectrometer (PCS) (Brookhaven, BI-90 plus, Holtsville, NY, USA). A 10-mg not dextran nanoparticle was dispersed in 5 ml of isopropyl alcohol and used for PCS analysis. Encapsulation efficiency of proteins and recovery of dextran nanoparticle The encapsulation efficiency of dextran nanoparticles was determined as follows: the amount of BSA, GM-CSF, and MYO recovered from the dextran nanoparticle was determined by the Micro BCA kit. The dextran nanoparticles loaded with proteins obtained were weighed and then dissolved in deionized water for Micro BCA determination. All measurements were performed in triplicate. The encapsulation efficiency of protein and recovery of the dextran nanoparticle were calculated as follows: (1) (2) Assay of protein aggregation The BSA, GM-CSF, and G-CSF were selected as model proteins to examine the protein aggregation during the preparation process. The size-exclusion chromatography-high- performance chromatography (SEC-HPLC) was used to identify proteins and analyze the monomer protein content recovered. SEC-HPLC provides information on the size of the proteins and the presence of aggregated proteins.