Figure 3.

TPCs Are Necessary for TCR-Triggered Ca²⁺ Release via NAADP

(A–C) Ca²⁺ oscillations in Jurkat 1G4 cells induced by 2.5–10 μM NAADP/AM were inhibited by 10 μM trans-Ned-19 or 1 μM bafilomycin A1. Single-cell Ca²⁺ traces (A) and summary of maximum peak Ca²⁺ changes and MER (B and C) are shown. n = 88–103 cells.

(D–I) Jurkat 1G4 cells were nucleofected with 100 pmol siRNA (nontargeting [NT], TPC1 and TPC2) or without siRNA (Mock) and incubated for 72 hr (D–G) or 48–72 hr (H and I). siRNA-treated cells were stimulated with 10 μM NAADP/AM (D–F) or by contact with 10 μg/ml anti-human CD3-coated coverslips (G–I). Single-cell Ca²⁺ traces (D and G) and summary of peak Ca²⁺ changes (E–F and H–I) are shown for n = 213–383 cells from six independent siRNA experiments.

Error bars are mean ± SEM. See also Figure S3.