Figure 6

Genomic organization and evolutionary relationship of nucleolin and NRPs in different eutherian mammals. A) Schematic representation of genomic organization of nucleolin on (chromosome 9) and NRP gene on (chromosome 15). Exons are represented in grey bars interrupted with white bars as introns. The black region of 3^rd exon and complete exon 4 of nucleolin are absent in the NRP gene and the arrow represents the corresponding exons of nucleolin found in NRP. The colored bars in the upstream sequences represent the different transcription factor binding sites. B) Conservation of the NRP across 8 different mammals. ClustalW was used to generate the dendrogram. C) Domain alignment of Nucleolin and NRP from different species showing the missing stretch of acidic amino acids in mouse and rat NRP as against nucleolin of other species. D) Multiple tissue northern blot analysis using rat polyA RNA blot. Upper panel, hybridisation with a probe specific to nucleolin spanning the exon 4. Bottom panel, hybridisation with full length nucleolin common to both. E) Colocalisation of nucleolin and NRP. pCMV myc-NRP was transfected into GC1-spg cells. Anti Myc was used to detect NRP in the transfected cells, while the endogenous nucleolin was detected by polyclonal nucleolin antibody followed by appropriate secondary. Panel (a), nucleolin; panel (b), myc; and panel (c), merge of a and b.