Figure 2. WAVE1 regulates ROS production and Ca²⁺ homeostasis in apoptosis.

(A) Knockdown WAVE1 by siRNA in Jurkat or HL-60 cells and treated with ADM (1 μg/ml), thapsigargin (100 nM),or H₂O₂ (250 μmol) for 24 h, and ROS production, ER Ca²⁺ release (cytosolic free calcium) and mitochondria Ca²⁺ load was analyzed by flow cytometry (n=3, * P < 0.05 versus control siRNA group). UT, Untreated group, set as 1. (B) Knockdown WAVE1 cells were treated with thapsigargin (100 nM) or H₂O₂ (250 μmol) with or without FCCP (2.5 μmol) for 24 h, and mitochondria Ca²⁺ load was analyzed by flow cytometry (n=3, * P < 0.05). Untreated group, set as 1. (C) Knockdown of Bcl-2 by shRNA in WAVE1 overexpressin cells restored ROS and Ca²⁺ production. HL-60 cells transfected with various plasmids were stimulated with thapsigargin (100 nM) for 24 h, and cellular levels of ROS, cytosolic free calcium, and mitochondria Ca²⁺ load was analyzed by flow cytometry (n = 3, * P < 0.05).