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. 2009 May 21;284(28):18707–18714. doi: 10.1074/jbc.M109.017483

FIGURE 4.

FIGURE 4.

Phosphoacceptor Ser-5, -6, and -7 residues are essential for in vivo CAPS activity in evoked vesicle exocytosis. A, CAPS was down-regulated in PC12 cells by transfection of an short hairpin RNA plasmid (lower panel) resulting in a 5-fold reduction in the number of exocytic events evoked by 56 mm K+ depolarization (upper panel). GAPDH, glyceraldehyde-3-phosphate dehydrogenase; WT, wild type. B, transfection with a pcDNA3.1-CAPS-tag red fluorescent protein harboring silent mutations enabled re-expression of wild-type or S5A/S6A/S7A CAPS in down-regulated PC12 cells (lower panel). Wild-type CAPS expression restored evoked exocytosis, whereas S5A/S6A/S7A CAPS expression failed to do so (upper panel). Error bars indicate S.E. for n = 13–17 cells. AFM, arbitrary fluorescence units.