FIG. 2.
Phosphorylation of in vivo PKA substrates is increased by gpb1Δ gpb2Δ mutations. (A) Extracts made from the following strains expressing Sfl1p-myc from plasmid pXP181 (27) (provided by X. Pan and J. Heitman) were analyzed by SDS-PAGE and immunoblotting with anti-myc antibody 9E10: wild-type strain SKY762 (lane 1), gpb1Δ gpb2Δ strain HS182-3B (lane 2), tpk2Δ strain HS192-2C (lane 3), and gpb1Δ gpb2Δ tpk2Δ strain HS196-5A (lane 4). (B) Extracts made from the following strains expressing Msn2p-GFP fusion protein from plasmid pAdh1-Msn2-GFP (12) (provided by C. Schüller and W. Reiter) were analyzed by SDS-PAGE and immunoblotting with antibodies to P-CREB (S133) and GFP: wild-type strain SKY762 (lane 1), gpb1Δ gpb2Δ strain HS182-3B (lane 2), SKY762 carrying vector YCplac33 (lane 3), and SKY762 carrying plasmid YCp50-RAS2ala18val19 (lane 4).