The expression of GFP-BLM is inducible in stable transfected BS cells.
A. Equal amounts of total protein (10 μg) from whole cell lysates were resolved on a SDS-polyacrylamide gel and transferred to a membrane. The bound primary α-BLM and goat α-rabbit alkaline phosphatase conjugated secondary antibody complexes were quantitated using a ECF fluorescent alkaline phosphatase substrate. 1) GM0637G normal SV40-transformed fibroblast cells (0.17), 2) HG2522-1a cells + GFP-BLM gene no inducer (0.03), 3) +0.0625 μg/ml Tet (0.05), 4) +0.125 μg/ml Tet (0.11), 5) +0.25 μg/ml Tet (0.16), 6) +0.5 μg/ml Tet (0.3), 7) +1 μg/ml Tet (0.3), 8) +2 μg/ml Tet (0.14). The ECF fluorescent product was visualized directly with a Molecular Dynamics Storm PhosphorImager. B. The fluorescent image was quantitated using Imagequant. The amount of BLM in the lysates was normalized to the amount in equivalent total protein in the GM0637G cells. The data points are the average of two experiments. The standard deviations for each concentration are reported in parentheses above.
Yankiwski et al. BMC Cell Biology 2001 2:11 doi:10.1186/1471-2121-2-11