FIG. 13.

Far-Western analysis of the binding of GR_C460Y and GR_C460Y/L501P peptides to proteins extracted from early zebra fish embryos. (A) Protein lysates were prepared from early-blastula-stage and early-gastrula-stage dissected zebra fish embryos from which the chorion had been removed. A Coomassie blue-stained SDS–12% polyacrylamide gel analysis of 25 μg of each extract is shown. (B) Following transfer to nylon membranes, the extracts were denatured and renatured and the membranes were hybridized with either GR_C460Y (lanes 1 and 2) or GR_C460Y/L501P (lanes 3 and 4) peptides labeled with ³²P to the same specific activity at a PKA phosphorylation site added to the C termini of the peptides. Incubation, washing, and exposure to autoradiographic film of the membranes were performed identically for each probe.