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Fig. 4. Confirmation of the interaction between DRAL/FHL-2 and the N2B and is2 regions of titin by colocalisation and pull-down experiments. (A) Transient transfection assays. GFP-N2B (a) and DRAL/FHL-2-FLAG (a') are localised in a similar pattern in transiently transfected NRC, with a broad doublet flanking the Z-disc (arrowheads) and a weaker striation at the M-band. A similar colocalisation can be observed in neonatal rat cardiomyocytes transiently transfected with GFP-is2 (b) and DRAL/FHL-2-FLAG (b'). (B) Pull-down assays: GFP-N2B and GFP-is2 interact with DRAL/FHL-2-GST but not with GST alone. Proteins in the supernatant and pellet fractions were detected by immunoblotting using a GFP antibody.