Fig. 4.

Vacuolar localization of AtCaM15. (A) AtCaM15 fused with 3xFLAG tag at the N terminus (3xFLAG::AtCaM15) or C terminus AtCaM15::3xFLAG) was introduced into yeast W303 strain and the isolated vacuoles were incubated with proteinase K (ProK) in the presence or absence of Triton X-100 (TX) as indicated. The data are representative of three independent experiments. (B) Arabidopsis leaf protoplasts were transiently transformed with EGFP or AtCaM15::EGFP plasmids. Transformed protoplasts were stained with FM4-64 to visualize membranes, and images were acquired as described in Materials and Methods.