(A) Total protein extracts of WT, reg1Δ and snf1Δ cells were prepared before and after 2DG addition at the indicated concentrations for 10 min and immunoblotted using anti-phospho-AMPK and anti-polyHistidine antibodies. (B)
Left, Total protein extracts of WT, reg1Δ and snf1Δ cells were prepared before and after 2DG addition for the indicated time and immunoblotted using anti-phospho-AMPK and anti-polyHistidine antibodies. Right, quantification of the signals (adjusted p-value is indicated, one-way ANOVA). (C) Total protein extracts of WT and reg1Δ cells expressing Snf1-GFP and Rod1-Flag were prepared before and after 2DG addition for the indicated time and immunoblotted using anti-Flag, anti-phospho-AMPK and anti-GFP antibodies. (D) Total protein extracts of WT, reg1Δ and snf1Δ cells expressing Mig1-Flag were prepared before or after 2DG addition for 10’ and 2h and immunoblotted using anti-Flag antibodies.