Fig. 3. (A) Western blot analysis of the eight potential isoforms cloned as EGFP-fusion proteins. COS-7 cells were transfected with pEGFP-splice variant expression plasmids indicated above. 48 hours after transfection extracts of COS-7 cells were prepared and separated by SDS-PAGE; each lane contains 20 µg of total protein. (B) Western blot analysis of splice variant FL-3/7 as EGFP-fusion protein. i28, MCF7, RT112, EJ28, HeLa and COS-7 cells (indicated above) were transfected with pEGFP-splice variant FL-3/7 expression plasmid. 48 hours after transfection cell extracts were prepared and separated by SDS-PAGE; each lane contains 20 µg of total protein. The western blots were probed with GFP-antibodies followed by incubation with alkaline phosphatase conjugated secondary antibody, and visualised by NBT/BCIP. (FL, full length).
