Fig. 5
Essential role of Kr-h1 phosphorylation in the interaction with transcriptional cofactors. A Upper panel: immunoprecipitation (IP) and western blot (WB) showing the interaction of Flag-Kr-h1, Flag-Kr-h1S154A, or Flag-Kr-h1S154D with Flag-CtBP. Middle and lower panels: the expression of above recombinant proteins in S2 cells. α-Kr-h1, Kr-h1 antibody; α-Flag, Flag antibody. WT, wildtype; MT, mutant. B Luciferase reporter assays after co-transfection of pGL4.10-4×E93-623 to -606 and pAc5.1/Flag-CtBP plus pAc5.1/Flag-Kr-h1, pAc5.1/Flag-Kr-h1S14A, or pAc5.1/Flag-Kr-h1S154D into S2 cells. Co-transfection of pGL4.10-4×E93-623 to -606 and pAc5.1/Flag-Kr-h1 was used as the control. Methoprene was applied at 10 μM. Means labeled with different letters indicate significant difference at P<0.05. n=4. C Upper panel: IP and WB showing interaction of Flag-Kr-h1, Flag-Kr-h1S154A or Flag-Kr-h1S154D with Flag-CBP. Mid and lower panels: the expression of above recombinant proteins in S2 cells. α-Kr-h1, Kr-h1 antibody; α-Flag, Flag antibody. WT, wildtype; MT, mutant. D Luciferase reporter assays after co-transfection of pGL4.10-4×RL36-1647 to -1632 and pAc5.1/Flag-CBP plus pAc5.1/Flag-Kr-h1, pAc5.1/Flag-Kr-h1S154A or pAc5.1/Flag-Kr-h1S154D into S2 cells. Co-transfection of pGL4.10-4×RL36-1647 to -1632 and pAc5.1/Flag-Kr-h1 was used as the control. Methoprene was applied at 10 μM. Means labeled with different letters indicate significant difference at P<0.05. n=4