MTCL2 is preferentially enriched in the dendrites of cerebellar granule neurons. A, Cerebellar sections from 4-week-old mice are stained for calbindin and NeuN (left panel) or MTCL2 and α-tubulin (middle and right panels). Scale bars, 250 µm. B, Representative images of isolated CGNs cultured for 5 d and stained for acetylated tubulin, MTCL2, and MAP2. Scale bar, 20 µm. C, Correlation between the immunofluorescence intensities of MTCL2 and MAP2 in acetylated tubulin-positive neurites. A total of 100 neurites from three independent experiments were analyzed. D, Representative images of CGNs at DIV1–DIV5 stained for tau1, MTCL2, and MAP2. In the merged view, nucleus staining is included to clarify the positions of cell bodies. The arrowheads indicate tau1-positive neurites exhibiting MTCL2 signals in immature CGNs. Scale bar: 20 µm. E, Time-dependent increase in the relative intensities of the MTCL2 (solid line) and MAP2 (dotted line) signals in dendrites. Neurites exhibiting MAP2 signal intensities three times greater than the background intensity were analyzed as dendrites. Data represent the means ± SD of the average values for each of the three independent experiments (more than 67 neurites were analyzed for each DIV stage). Welch’s two-sample t test was used to examine statistical significance. F, Western blot analysis of MTCL2 and MAP2 expression in primary cultured CGNs from various DIVs. A representative result of two independent experiments is presented.