FIG. 1.

Reduced formation of astrocytes in the developing cortex of Bax-KO mouse. Cerebral hemispheres were dissected from WT and Bax-KO mice at P2 and P14 and were subjected to immunoblot analyses for expression of astrocytic markers GFAP and S100β and neuronal markers TuJ1 and MAP2. (a) Representative immunoblots of WT and Bax-KO cortices at P2 are shown. (b) Each GFAP and MAP2 band of WT and Bax-KO animals was scanned for quantification. The data were obtained from the P2 offspring (n = 4 for Bax-KO and n = 9 for WT) from three pregnant mice and from P14 offspring (n = 3 for Bax-KO and n = 5 for WT) from two pregnant animals. *, significantly different from WT control at a P value of <0.001. (c and d) Reduction in the number of GFAP-positive astrocytes in Bax-KO brain. Brain cell suspensions from P10 mice were fixed, permeabilized, and immunostained with anti-GFAP antibody (Ab). The percentage of GFAP-labeled cells was obtained by a flow cytometer. Shown in panel c are representative profiles of the flow cytometry analyses. The boxed area includes the GFAP-labeled cells recognized and separated on the basis of secondary antibody fluorescence emission pattern. SSC, side scatter. The number of astrocytes per brain, as shown in panel d, was calculated by multiplying the astrocyte ratio by the total cell number in the brain. Means ± SEM of six (WT) or four (Bax-KO) animals were from three independent experiments. *, P < 0.05 in Student's t test comparison.