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文献和实验for cell lysis: 10 mM Tris-HCl, pH 7.4,50 mM NaCl, 5 mM EDTA, 50 mM NaF, 30 mM Na 4 P 2 O 7 ,and 1% (v/v) Triton-X100. 5. Protease inhibitors and phosphatase inhibitor are added fresheach time with the following final concentrations: 10 μg/mLaprotinin
: L-Wnt3A (ATCC , Manassas, VA; ATCC# CRL-2647) (see Note 1 ). 2. Cell culture medium: Dulbecco’s minimum essential medium(DMEM) supplemented with 10% (v/v) fetal bovine serum(FBS), 1:100 dilution of penicillin–streptomycin solution with10,000 U
transfer vessels. Each day students will observe these transfer cultures with an inverted phase contrast microscope, and remove aliquots for cell counting with a hemacytometer. Simultaneously, they will check on the viability of the cells through a dye
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