Fig. 3. Modulation of gene expression by FGF9. (A) Differential hybridization to Atlas rat gene array. Total RNA prepared from either RCS cells incubated with FGF9 and heparin for three hours or untreated cells was used to generate two radiolabeled cDNA probes. The probes were hybridized to the Clontech Atlas membranes (7738-1) as described in Materials and Methods. Each cDNA is present on the filter in two adjacent spots. The frames indicate several genes regulated by FGFR3. The previously unreported, upregulated genes were (fold-induction is given in brackets) c-Jun (x2.5), Jun D (x15.1), Fra 2 (x5.6), cyclin D1 (x2.3), NF-B1(p50/p105) (x3.6), STAT3 (x3.1) and Ezrin (x4.0), whereas ID1 was downregulated three-fold. (B) Kinetics of protein expression following FGF9-induced growth arrest. RCS cells were exposed to FGF9 and heparin for the indicated time intervals, lysed and analyzed by SDS-PAGE and western blotting with anti c-Jun, Jun-D, p21, Id1 and Ezrin antibodies.

(C) Immunohistochemical analysis of epiphyseal growth plates from normal and transgenic G380R mutant h-FGFR3 expressing mice. Immunostaining for Rel A (NF-B p65) was performed on sections of proximal tibia growth plates from normal and transgenic littermates harboring the G380R mutated FGFR3. The different zones of the growth plate (PZ-proliferation zone, MZ-maturation zone, HZ-hypertrophic zone) are noted.

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