Fusion protein expression cassette design and protein expression

(A). Each expression cassette has GFP at the N-terminus with the respective actin binding protein at the C-terminus. Gene expression is driven by the actin 15 promoter. The numbers above each protein fragment represent the amino acids included in the construct. (B) Western Blot analysis of whole cell lysates of cells expressing constructs. Total cellular protein from each cell line was separated by SDS-PAGE, blotted onto PVDF and each panel was probed with an antibody specific for filamin, α-actinin or GFP. abpA/abpCcell line; lane 1, 5. AX2 wt; lanes 3, 6, 10. GFP-FLN; lanes 2, 11. GFP-FLNABD; lanes 4, 12. GFP-α A; lanes 7, 13. GFP-α AABD; lanes 8,9,14. pDNeoGFP base vector: lane 15. Lane 9 is loaded with twice the amount of extract as lane 8 in order to more easily visualize the bands.Copyright information:Taken from "Actin binding domains direct actin-binding proteins to different cytoskeletal locations"http://www.biomedcentral.com/1471-2121/9/10BMC Cell Biology 2008;9():10-10.Published online 13 Feb 2008PMCID:PMC2275727.