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Fig. 3. FL-dextran distribution in electroporated cells. (A-G) HeLa cells. Passive diffusion rates of 19.5 kDa FL-dextran. Cells were electroporated with 19.5 kDa FL-dextran and nucleoplasmic to cytoplasmic fluorescence ratio (Fnp/Fcytoplasm) was monitored for 60 min in 2 mM EGTA (
) or 2.1 mM Ca2+ containing electroporation buffer (
). (B) Graphs show Fnp/Fcytoplasm ratios of cells in the specified conditions. Order of groups of columns from left to right. Cells in 2 mM EGTA for 15 minutes; cells in 2 mM EGTA and 20 µg/ml WGA for 15 minutes; addition of 2 mM Ca2+ for 1 hour; addition of 2 mM Ca2+ for 1 hour with 20 µg/ml WGA electroporated; addition of 10 µM BAPTA-AM before electroporation in 2 mM EGTA. (C-G) Confocal images of 4.3-, 9.5-, 19.5-, 40- and 77-kDa FL-dextrans, respectively, in HeLa cells in 2 mM EGTA imaged 15 minutes after electroporation. (H,I) Dextran distributions in cortical glial cells (H) and cortical neurons (I). Order of groups of columns from left to right: cells in 2 mM EGTA for 15 minutes; cells in 2 mM EGTA and 20 µg/ml WGA for 15 minutes; addition of 2 mM Ca2+ for 1 hour; addition of 2 mM Ca2+ for 1 hour with 20 µg/ml WGA electroporated.
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