Figure 1.

Knockdown of Orai1 reduces TG-induced SOCE in RAW264.7 cells. A) RNA was isolated from RAW cells infected with lentivirus expressing either scrambled or Orai1 shRNA for 5 d. Expression levels of mouse Orai1 message were measured by qRT-PCR. Statistical significance was evaluated by Student's t test. Data represent 2 independent experiments with similar results. Error bars denote se between samples performed in triplicate. B) Ca²⁺ imaging of RAW cells infected with the scrambled or Orai1 shRNA lentivirus. Cells were loaded with 5 μM fura-2-AM dye, and SOCE was revealed by readdition of 1.8 mM Ca²⁺ after TG-induced Ca²⁺ release from ER and measured by optical imaging of intracellular Ca²⁺ concentration. C, D) Peak value (C) and initial rates (D) of SOCE increase from B were measured. Statistical significance was evaluated by Student's t test. All data are means ± se from 3 independent experiments (30–40 cells/experiment analyzed). *P < 0.05, ***P < 0.001.