Figure 5.

The distribution of SPK2 in the cytoplasm and nuclear fraction of neural cells. (a) LV-vector-HT22 and LV-SPK2-HT22 cells were fixed with 4% paraformaldehyde and processed for immunofluorescence. Representative images were stained with DAPI (blue) and antibody against SPK2 (red). Scale bar=10 μm. (b) Nuclei and cytoplasm of LV-vector-HT22 and LV-SPK2-HT22 cells were extracted and subjected to western blot analysis. (c) Cortical neurons were infected with LV-SPK2 or LV-vector at DIV2. At DIV 7, the nuclei and cytoplasm of neurons were extracted and subjected to western blot analysis. (d) The distribution of SPK2 in the cytoplasm and nuclear fraction of neurons after isoflurane preconditioning (ISO). The neurons were exposed to 2% ISO for 30 min. Twenty-four hours later, the cytoplasmic and nuclear fractions of neurons were extracted and subjected to western blot analysis. (e) ISO increased the colocalization of SPK2 and Bcl-2 in the cytoplasm of neurons. Neurons were fixed with 4% paraformaldehyde and processed for immunofluorescence. Representative images were stained with DAPI (blue), and antibodies against SPK2 (red) and Bcl-2 (green). Scare bar =10 μm. n=3 independent experiments