Fig. 6. Overexpression of TLS-GFP in TLS-null neurons increases amount of Nd1-L mRNA in dendrites. Primary cultures of TLS-null hippocampal neurons (21 DIV) were infected with adenovirus containing TLS-GFP to overexpress TLS protein. TLS-GFP-expressing TLS-null neurons were then subjected to cell in situ hybridization using an Nd1-L-specific probe. (A) TLS-null neurons express hardly any Nd1-L mRNA in their dendrites. Bar, 10 µm. (B) A quantitative representation of A. Overexpression of TLS-GFP increases the amount of Nd1-L mRNA by fivefold compared with that observed in TLS-null dendrites (P<0.05, n=50 individual dendrites). (C) Immunostaining with anti-GFP antibody after cell in situ hybridization for Nd1-L mRNA. TLS-GFP was colocalized with Nd1-L transcripts in mature spines of wild-type neurons (top). In TLS-null neurons expressing TLS-GFP, Nd1-L transcripts were also found in spines together with TLS-GFP (bottom, arrows). Bar, 30 µm.