Figure 4From: The subcellular localization of IGFBP5 affects its cell growth and migration functions in breast cancer Western blot analysis of IGFBP5 expression in both cell lysate and conditioned medium. One million cells were cultured in 100-mm diameter tissue culture dish in the complete medium for 24 h followed by an additional 24-h incubation in the serum-free medium. Both cell lysate and conditioned medium were collected. The conditioned medium was concentrated 3 times using YM-10 column (Millipore) and 40 μg of protein from the cell lysate and 45 μl of the concentrated conditioned medium were subject for western blot analysis. The blot was re-probed with anti-actin to normalize for protein loading. wt, wild-type and mt, mutant form IGFBP5.Back to article page