2.4. siRNA Knockdown

RD cells were seeded at 0.25 × 10⁶ per well in 6-well plates, and after 24 h, growth media was replaced with 2 mL Accell delivery media supplemented with 2% FBS and 2 μM siRNA against ezrin, radixin, moesin, or a combination of all three (Dharmacon, Lafayette, CO, USA). 2 μM nontargeting scrambled siRNA was used as a negative control. After 48 h of incubation, 1 mL of fresh delivery media with 2 μM siRNA was added to each well. At 96 h, viable cell counts were determined by trypan blue, and cells were lysed for Western blotting.