Figure 4.

Effect of Highly Purified ANN33/35 Preparation on [Ca²⁺]_cyt of Protoplasts from Mature Root Epidermal Cells.

(A) Example of an annexin-induced increase in [Ca²⁺]_cyt. Highly purified maize annexin preparation (5 mg/L) added (arrow; closed circles) to the extracellular membrane face of Arabidopsis root epidermal protoplasts (constitutively expressing aequorin) in 10 mM external Ca²⁺ caused a transient increase in [Ca²⁺]_cyt. The control experiment adding buffer alone is also shown (open circle).

(B) Mean ± se [Ca²⁺]_cyt response to 5 mg/L Zm ANN33/35 (closed circle; n = 3) or buffer alone (open circle; n = 4).

(C) Mean ± se [Ca²⁺]_cyt response to 5 mg/L Zm ANN33/35 preparation (closed circle; n = 4) or buffer alone (open circle; n = 4) both in the presence of 100 μM verapamil (Vp).

(D) Mean ± se [Ca²⁺]_cyt response to 5 mg/L Zm ANN33/35 (closed circle; n = 4) or buffer alone (open circle; n = 4) both in the presence of 300 μM Gd³⁺.

(E) Mean ± se [Ca²⁺]_cyt response to 5 mg/L immunoprecipitated Zm ANN33/35 (IP, closed circle; n = 3) or buffer containing equivalent anti-Zm ANN33/35 antibody (open circle; n = 3).

(F) Mean ± se peak [Ca²⁺]_cyt responses from (B) to (E) and the mean response to preimmune serum at an equivalent concentration to antibody (Ab) used in (E) (n = 3).