We washed and harvested 4 × 107 MIN6 cells, stably expressing the G264S mutant, in Hank's balanced salt solution, pelleted cells by centrifugation (300g, 5 min), and resuspended in washing solution (30 mM KCl, 140 mM NaCl, 10 mM EDTA, 25 mM Tris–HCl (pH 7.4)). After repeating...
Immunofluorescence Assays—For immunolocalization assays, 15-DAP immature maize embryos were fixed in an ethanol/formaldehyde/glacial acetic acid (80:3.5:5) fixative solution for 1 h at room temperature and for 7 days at 4 °C. Embryos were dehydrated and embedded in paraffin. 8-μm sections ...
A total of 20 μL of CellTiter 96 Aqueous One Solution Cell Proliferation Assay (MTS; 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) (Promega) was added to react for 3 hours. Measurement of absorbance of the samples at 490 nm ...