Fig. 2.

EGCG induces phosphorylation of TbACC. BF and PF ACC-myc lysates were treated with 100 μM EGCG or DMSO as solvent control in the presence of HALT phosphatase inhbitor cocktail. (A) BF and (B) PF TbACC-myc immunoprecipitates (∼5 × 10⁸ cell equivalents/lane) were resolved by SDS–PAGE and stained with Pro Q Diamond phosphoprotein gel stain to detect phosphorylated ACC (ACC-p; upper panels). Identically loaded gels were blotted to nitrocellulose and probed for total ACC with SA-HRP (ACC-t; bottom panels). Representative gels and blots are shown. Unlabeled upper band in panel A is an unknown phosphorylated contaminant. (C) Densitometric quantitation of phosphorylated ACC from phospho-stained gels normalized to total ACC detected by SA-HRP blotting. The mean of 3 independent experiments is shown. Error bars indicate the ±SEM. The * indicates P < 0.05 and ** indicates P < 0.00001 for the difference between DMSO control and EGCG-treated conditions (student’s t-test).