Figure 2

cDNA and siRNA transfection of hard-totransfect cells. (a) Western blot of differentiated HL-60 cells electroporated with siRNAs to knock down lamin A/C. (b) Lamin A/C immunostaining of HL-60 differentiated cells treated with LMNA-targeting (left) and non-targeting (middle) siRNA. Scale bars, 200 μm. Fluorescence intensity distributions of lamin A/C staining revealed 67% knockdown (right). (c) Differentiated HL-60 cells electroporated with eGFP cDNA and plated on a fibronectin-coated coverslip. Scale bar, 20 μm. (d) Under-agarose chemotaxis assay of GFP-electroporated HL-60 cells accumulating at the source of the chemoattractant fMLP (red circle) 6 h after exposure to the gradient (left), and enlarged views of the cells before (top right) and after (bottom right) initiation of chemotaxis. Scale bars, 1 mm and 200 μm (close-ups). (e) Rat primary hippocampal neurons 3 d after electroporation of GFP cDNA. Scale bar, 150 μm. (f–h) Differentiated 3T3-L1 adipocytes (f; left, fluorescence micrograph; right, overlay of transmitted light and fluorescence images), HUVEC (g) and RBL-2H3 cells (h) electroporated with eGFP cDNA. Scale bars, 20 μm (f (right), g and h) and 50 μm (f;left).

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