HeLa Kyoto cell cultures were imaged 24–48 h after the transient lipofectamine transfection before and immediately after 2.5 µM Ionomycin addition using a laser spinning-disk Andor XDi Technology Revolution multi-point confocal system (Andor Technology, Belfast, UK) as previously described [9 (link)].
For FRAP experiments, HeLa Kyoto cells were treated as described above and imaged using a Leica SP5 STED confocal microscope (Leica-Microsystems, Bensheim, Germany) and 70% of 488 nm laser power for bleaching (power at 100%—65 mW) during 1000 ms, with the capture settings: 100 ms per frame, 20 and 600 frames before and after bleaching, respectively, resolution—16 × 16 pixels, pixel size—0.38 µm.
For FRAP experiments, HeLa Kyoto cells were treated as described above and imaged using a Leica SP5 STED confocal microscope (Leica-Microsystems, Bensheim, Germany) and 70% of 488 nm laser power for bleaching (power at 100%—65 mW) during 1000 ms, with the capture settings: 100 ms per frame, 20 and 600 frames before and after bleaching, respectively, resolution—16 × 16 pixels, pixel size—0.38 µm.
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