Phosphocofilin intensity decrease upon LIMK inhibition.

To reduce the spot crowding around the phosphocofilin spot, a linear 4–8 pH gradient was used. This does not allow the visualization of the basic cofilin spot (spot I), and brings the median spot (II) to the border of the gel, rendering its quantitative measurement unreliable. By contrast, the phosphocofilin spot (III) is more separated from neighboring spots and can be more precisely quantitatively measured. It is also worth noting that spot IV, which is not detected by the anti phospho S3 peptide antibody, does not respond to Pyr-1 treatment. In these images, proteins were detected by silver staining.