3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2 h tetrazolium bromide mtt

SW480 cells were seeded into 96-well plates at a density of 8 × 10³ cells/well. After incubation at 37 C overnight, cells were treated with 0.4, 0.8, 2, 3 and 6 mM of crocin (catalog number: B21336; Shanghai Yuanye Biological Technology Co., Ltd., Shanghai, China) for 24 h at 37 C. Thereafter, 5 μl of 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) (Shanghai Yuanye Biological Technology Co., Ltd., Shanghai, China) was added to each well, to obtain a final concentration of 5 mg/ml MTT. The plates were then incubated at 37 C for a further 4 h, at which point the supernatant was aspirated, 100 μl of dimethyl sulfoxide (catalog number: DH105-2; Beijing Dingguo Changsheng Biotechnology Co., Ltd., Beijing, China) was added to each well, and finally the absorbance of the plates was measured at 490 nm using a Synergy™ 4 Microplate Reader (Biotek, Winooski, Vermont, United States).

Branched polyethylenimine (bPEI, 25 kDa) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Methotrexate (MTX) and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) were purchased from Shanghai Yuanye Biological Technology (Shanghai, China). mPEG-NH₂ (2000 Da) was purchased from Yarebio (Shanghai, China). Linolyl chloride (LC) was obtained from Tokyo Chemical Industry Co., Ltd. (Shanghai, China). Survivin siRNA: Sense (5′–3′): mGCAGGUUCCUmUAUCUGUCAdTdT; Antisense (5′–3′): UGAmCAGAmUAAGGAACCUGmCdTdT; Survivin siRNA negative control: Sense (5′–3′): mUUCUCCGAACmGUGUCACGUdTdT; Antisense (5′–3′): ACGmUGACmACGUUCGGAGAmAdTdT, and Cy3, 5′-FAM and Cy5-labeled survivin siRNA for cellular uptake and biodistribution studies were synthesized by Ribo Biochemistry (Guangzhou, China). HeLa cells were purchased from ATCC (Rockefeller, MD, USA). 4′,6-Diamidino-2-phenylindole (DAPI) and Lyso Tracker™ Green DND-99 were purchased from Invitrogen Co. (Carlsbad, CA, USA). The Dihydrofolate Reductase Assay Kit was also purchased from BioVision (S. Milpitas Blvd., Milpitas, CA, USA). All chemical reagents used were of analytical grade.

PU (pellethane 2363-80AE) was provided by Lubrizol Corporation (Wickliffe, OH, USA). Methylenediphenyl 4,4′-Diisocyanate (MDI) was bought from Aladdin Chemical Co. Ltd. (Shanghai, China). N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), toluene, and triethylamine were purchased from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China), and HA was obtained from Xianding Biotechnology Co. Ltd. (Shanghai, China). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC∙HCl), N-Hydroxysuccinimide (NHS), Lysozyme (LYS), bovine fibrinogen (BFG), human serum albumin (HSA), and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) were obtained from Yuanye Bio-Technology Co. Ltd. (Shanghai, China). Sodium dodecyl sulfate (SDS) and the Micro BCA Protein Assay Kit were obtained from Sangon Biotech (Shanghai, China) Co. Ltd. Fetal bovine serum (FBS) and Dulbecco’s modified Eagle medium (DMEM) medium were obtained from Gibco (Grand Island, NY, USA). E. coli (DH5 alpha), S. aureus (ATCC 6538), and P. aeruginosa (ATCC 2785) and lysogenic broth (LB) medium were provided by Professor Cui, East China University of Science and Technology (ECUST, Shanghai, China). PHMG was provided by Professor Guan, ECUST.