Figure 3.

Comparisons of the signal intensities of miRNA ISH in fresh and fixed brain sections with or without proteinase K digestion. In situ detection of miR-34a (A) and miR-181a (B) was performed in adult mouse brain sections. The hybridization temperature was 37°C below the T_m of each RNA. Detection was carried out using the NBT/BCIP colorimetric method. The sections were treated with or without proteinase K at the indicated concentrations. (A) Representative images of miR-34a in fresh (upper panels) and PFA perfusion-fixed (lower panels) brain sections are shown. The insert images are a higher magnification of the boxed brain sections including the TRN. Scale bars in (A), 1 mm; 0.2 mm in the inserted images. (B) ISH signals for miR-181a in fresh (left panels) and PFA perfusion-fixed (right panels) brain sections are shown. The insert images are a higher magnification of the boxed brain sections including the hippocampal DG (B). Scale bars in (B), 500 μm; 100 μm in the inserted images.