Fig. 2

Chromosomal localization of the PARs by FISH

(A) Human thrombin receptor maps to 5q13. FITC-labeled probe from a P1 clone containing the human thrombin receptor gene (see Materials and Methods) was hybridized to metaphase chromosomes. Specific hybridization at 5q13 is demonstrated (green). The FLpter (fractional length from the p terminus) was 0.455 ± 0.005. (B) Human PAR2 maps to 5q13. FITC-labeled probe derived from a P1 clone containing the human PAR2 gene (see Materials and Methods) was hybridized to metaphase chromosomes. Specific hybridization at 5q13 is demonstrated (green). The FLpter was 0.453 ± 0.006. (C) Colocalization of human thrombin receptor and PAR2 on interphase nuclei by two-color FISH. Probes for human thrombin receptor and PAR2 were simultaneously hybridized to interphase nuclei and detected by FITC or rhodamine fluorescence, respectively. Two single yellow spots are visible demonstrating colocalization of the probes on interphase chromosomes. (D) Colocalization of human thrombin receptor and PAR2 by FISH on “stretch” DNA by two-color FISH. Probes for human thrombin receptor and PAR2 were labelled with FITC and rhodamine, respectively, and simultaneously hybridized to chromatin fibers that were released from nuclei and stretched out prior to hybridization (see Materials and Methods). The areas of genomic DNA hybridizing to the thrombin receptor (green) and PAR2 (red) probes were distinct but adjacent.