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. 2011 May 16;193(4):769–784. doi: 10.1083/jcb.201008050

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© 2011 Chen et al.

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Figure 3. — Sarm1 is widely expressed in rodent brain and neurons. (A) Immunoblot of Sarm1 in different mouse organs. GAPDH is used as an internal control. (B) Regional distribution of Sarm1 in mouse brain. Cx, cerebral cortex; Hi, hippocampus; St, striatum; Th, thalamus; Cb, cerebellum; BS, brain stem. α-Tubulin was used as an internal control. (C) Staining patterns of Sarm1 in mouse brain. The top right shows the merged image of the MAP2/Sarm1 double stain in the CA1 region of the hippocampus. The top left and bottom panels depict the Sarm1 patterns in brain regions including layer five of the somatosensory cortex (Cx), the posterior thalamic nuclear group (Th), and the caudate putamen of the striatum (St). 2-mo-old mice were used in A–C. (D) Developmental expression profile of Sarm1. The plotted relative Sarm1 protein expression levels were obtained by normalization to the corresponding α-tubulin protein amounts. The results are the means of three independent experiments. Error bars indicate SEM. (E) Distribution of Sarm1 protein in biochemical subcellular fractions of adult mouse brain. H, total homogenate; P1, nuclei and cell debris; S1, supernatant of P1; P2, crude synaptosomal fraction; S2, supernatant of P2; LP1, lysed synaptosomal membrane; LS1, supernatant of LP1; P3, light membrane fraction; S3, soluble cytosolic fraction. PSD-95 enriched in the P2 and LP1 fractions was used as a quality control of fraction preparation. Molecular mass standards (kD) are indicated next to the gel blots. (F) Distribution of PSD-95 (red) and Sarm1 (green) in cultured hippocampal neurons at 21 DIV. Representative high-magnification images are shown on the top right. Arrowheads indicate the Sarm1 puncta overlapping with PSD-95; arrows indicate the Sarm1 puncta adjacent to PSD-95 puncta. The percentage of overlapped Sarm1 and PSD-95 is shown on the bottom right. The original images and the overlays shifted for 1 and 1.66 µm were analyzed. Error bars indicate mean values ± SEM. **, P < 0.01; ***, P < 0.005. Bars: (C) 30 µm; (F, left) 20 µm (F, top right) 2 µm.