Figure 6

Ccl19 is abnormally expressed in S8 hippocampus and is duplicated. (a) A northern blot of total RNA from spleen and hippocampus after hybridization with a Ccl19-specific probe is shown. An abnormally large Ccl19 transcript is detected only in the hippocampus of S8 mice, while the spleen of S8, S10 and SR1 all show normal Ccl19 expression. (b) Predicted amino-acid sequence of CCL19 from the hippocampal cDNA sequence of S8 mice reveals a putative truncation at the amino terminus of the protein relative to that of S8 spleen, and SR1 spleen and hippocampus. Arrows indicate the sites of mutations in the protein found in the S8 hippocampus. The region of the protein deleted by the mutations is highlighted in yellow. A box surrounds the first two conserved cysteines, which are adjacent and conserved in all β-chemokines. Colored boxes indicate the location of the signal peptide and the SCY-domain. (c) A Southern blot of S8, S10 and SR1 genomic DNA digested with EcoRI. The upper panel shows the signal from hybridization with a Ccl19 probe, and the lower panel shows the signal of the same blot hybridized with a control probe demonstrating relative DNA loading. The average Ccl19 signal intensity of each S8 lane is 1.9-fold greater than in SR1 and S10 when normalized to the control probe (p < 0.005 with a two-tailed Student's t-test). (d) A Southern blot of genomic DNA digested with BamHI. In this case, the lower panel shows the signal from hybridization with a Ccl19 probe, and the upper panel shows the signal of the same blot hybridized with a control probe demonstrating relative DNA loading. The average Ccl19 signal intensity seen in S8 is 2.6-fold greater than in SR1 and S10 when normalized to the control probe (p < 0.003 with a two-tailed Student's t-test).