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Figure 7


Fig. 7. p2 MEC responses to Fgf. (A) pMAPK and pAkt responses of Fgfr1{Delta}Frs/{Delta}Frs and control MECs stimulated with Fgf, assayed over a dose titration (Ai, 5 minute stimulations) and a 1 hour timecourse (Aii). RasGAP, loading control. (B,C) Crk and Frs2 activation responses to Fgf (50 ng/ml aFgf, 5 µg/ml heparin, 5'), assayed by IP-phosphotyrosine (4G10) western blot. Blots were stripped and reprobed with anti-Crk or anti-Frs2 (lower panels). *No antibody IP control. (D) Fgfr2 responses to Fgf (50 ng/ml aFgf, 5 µg/ml heparin, 5'), assayed by 4G10 IP-Fgfr2 western blot (upper panel). Total cell lysate Fgfr2 (middle panel) and actin (loading control, lower panel) western blots show relative protein levels. Matching arrowheads indicate bands of the same size recognized by different antibodies within B-D. (E) Relative Fgfr2 mRNA levels in homozygous embryos, determined by semi-quantitative RT-PCR. (F) Growth curves of p2 MECs derived from four mutant and two wild-type embryos cultured in DMEM, 4% FBS, 50 ng/ml aFgf.