The cells were removed from the dishes with 1.2 ml of phosphate buffer, twice washed, and incubated for 10 min on ice in the modified lysis buffer containing 50 mM Tris-HCl, pH 7.5, 0.5% Igepal CA-630, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 1 mM PMSF, 0.1 mM sodium orthovanadate and aprotinin, leupeptin, pepstatin (1 μg/mL each) as described earlier (Scherbakov et al., 2006). The protein content was determined by the Bradford method.