Anti c myc 9e10

The brain frozen sections or cultured neurons fixed in 4% paraformaldehyde for 15 min were incubated with pre-hybridization buffer (50% formamide [Wako], 5 × saline-sodium citrate buffer [SSC], 0.05% heparin sodium [Wako], 0.02% ribonucleic acid from torula yeast [Sigma]) for 60 min at 65 °C and then hybridized with a fluorescently labeled locked nucleic acid (LNA) probes, PolyT(25)Vn 5′-fluorescein (Exiqon 300510-04), Scramble-ISH 5′-fluorescein (Exiqon 300,514–04), and 5′-FAM-AGGCCAGGTCTTCTTCAGAAATCA-3′ (for exogenous FUS mRNA) (Exiqon) diluted to final concentration of 80 nM, for 24 h at 65 °C in a dark, humidified chamber. Next, sections were washed with 2 × SSC/formamide for 1 h at 65 °C and washed with TTBS (0.5 M NaCl, 0.02 M Tris HCl [pH 8.0], 0.1% tween 20) three times. Then immunohistochemistry was performed using anti-c-Myc 9E10 (1:1000), and anti-V5 (Cell Signaling, #06-549, 1:1000). Fluorescence intensity was determined by densitometry using ImageJ.

The brain frozen sections or cultured neurons fixed with − 20 °C methanol for 10 min were incubated with 500 nM SYTO RNASelect (Life technologies) for 20 min at room temperature as per the manufacturer's instruction. Then, immunohistochemistry was performed using anti-c-Myc 9E10 (1:1000) and anti-V5 (Cell Signaling, #06-549, 1:1000).