BDNF expression in the cerebellum and brain stem region.

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BDNF expression in the cerebellum and brain stem region. BDNF expression in the cerebellum and brain stem region. (A) mRNA levels of Bdnf and Nt-4 relative to those of Rpl13a in the cerebellum from wild-type (+/+) and Pex14ΔC/ΔC (ΔC/ΔC) BL/ICR mice at P3 were determined by real-time PCR (n = 4). (B) Sagittal sections of the cerebellum at P7 were stained with antibodies to GFAP (green) and BDNF (red). Merged views of the two different proteins and staining with Hoechst 33242 (blue) are shown. Scale bar, 20 μm. (C) In situ hybridization analysis of Bdnf mRNA in the brain stem region of wild-type (upper panels) and Pex14ΔC/ΔC (lower panels) mice at P0.5. Arrows indicate ION. Insets are higher magnification images of the dashed-line boxed regions. Scale bar, 100 μm. (D) Relative intensity of Bdnf mRNA staining by in situ hybridization shown in C was presented (n = 3). (E) Sagittal sections of the brain stem at P3 were stained with anti-BDNF antibody (red) and Hoechst 33242 (blue). Scale bar, 20 μm. (F) Fluorescent intensity of BDNF staining on the ION shown in E was quantified (n = 3). (G) The mRNA level of Bdnf in the brain stem region was quantified by real-time PCR (n = 3). (H) Brain stem lysates were analyzed by SDS–PAGE and immunoblotting using antibodies against BDNF and α-tubulin. The BDNF band was quantified (lower panel, n = 4). ns, not significant, *P < 0.05, **P < 0.01, by t test (A, D, F–H). Source data are available for this figure. Yuichi Abe et al. LSA 2018;1:e201800062 © 2018 Abe et al.