Immunofluorescence Detection of CCM2, MAP3K3, Phosphorylated MAP2K3/6 and Phosphorylated MAP2K4 during Mouse Preimplantation Development. Whole-mount indirect immunofluorescence of 1-cell, 2-cell, 4-cell, 8-cell, morula, and blastocyst stage mouse embryos with antisera raised against CCM2 (CCM2), MAP3K3 (MAP3K3), phosphorylated MAP2K3/MAP2K6 (P-MAP2K3/6) and phosphorylated MAP2K4 (P-MAP2K4). CCM2 and MAP3K3 immunofluorescence was distributed diffusely throughout the cytoplasm of each blastomere in all embryo stages. Phosphorylated MAP2K3/MAP2K6 (phosphorylated at Ser189/107) immunofluorescence was detected in the cytoplasm and nucleus of each blastomere in each embryo stage. Phosphorylated MAP2K4 (phosphorylated at Thr261) immunofluorescence was primarily localized to the nucleus of each blastomere in each embryo stage. In all confocal micrographs, green indicates positive staining for specific protein and red indicates F-actin (rhodamine-phalloidin). Scale bars represent 50 μm.
Fong et al. BMC Developmental Biology 2007 7:2 doi:10.1186/1471-213X-7-2