Fig. 3

Postnatal alcohol exposure (PAE) reduced the number of Purkinje neurons in the cerebellar vermis. Representative images of the cerebellar vermis stained for calbindin (red) to label Purkinje neurons and 4′,6-diamidino-2-phenylindole (DAPI, blue) to label cell nuclei. Purkinje cell number was quantified during the first withdrawal period on P4 (a–b), the third withdrawal period on P6 (c–d) and also on P45 (e–f) (as shown in Fig. 1a). The cerebellar vermis was divided into three lobule regions and quantified separately. Purkinje cells were counted in each image and the average cells falling within the counting frame per image was averaged. (**p < 0.01, ***p < 0.001, ****p < 0.0001). n = 4 animals from 4 litters. The thickness of the external and internal granule cell layers (EGL and IGL, respectively) was also quantified and results are shown in Additional file 3. Scale bars are 40 and 10 μm for low and high magnification images, respectively