Time-lapse quantification of FA dynamics after calpain 2 inhibition. (A) Western blot showing calpain 2 siRNA (sequence 1) efficiency in control and FLNa-silenced BT-20 cells. GAPDH was used as an internal control. MM, molecular mass. (B) Control (PSR) or FLNa-silenced (PSR-FLNa) BT-20 cells stably expressing EYFP-paxillin were transfected with calpain 2 siRNA, stimulated with EGF, and then analyzed by time-lapse spinning disk confocal microscopy. White arrows show the positions of paxillin-containing FAs. Four independent experiments were performed, and representative frames are shown. Bar, 5 µm. (C, left) Quantification of the persistence of EYFP-paxillin in control and FLNa knockdown cells. (right) Quantification of EYFP-paxillin disassembly. Quantifications show the mean ± SD from three independent experiments.