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FIGURE 3.
Subcellular localization of PIAS2b during early development. Staged albino oocytes and embryos were stained using PIAS2b primary antibody and secondary antibody conjugated to Alexa Fluor 568. A, PIAS2b is exclusively cytoplasmic at all stages of oogenesis. B, protein extract prepared from nuclei and enucleated oocytes (1.5 oocyte equivalents per stage) was analyzed by Western blotting using PIAS2b antiserum. Oocyte stages are indicated above each lane. Top panel, a blot of extract from enucleated oocytes. Bottom panel, extract prepared from isolated nuclei. Also shown is immunohistochemical staining of embryos at mid-blastula (C and D) and early gastrula (E and F) stages for PIAS2b. D, overlay of PIAS2b and TOPRO-3 nuclear staining of the midblastula embryo shown in C. Arrows point to individual nuclei. Images were collected at ×10 magnification unless indicated otherwise. |
