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. Author manuscript; available in PMC: 2014 Jun 1.
Published in final edited form as: Int J Biochem Cell Biol. 2013 Mar 1;45(6):1051–1063. doi: 10.1016/j.biocel.2013.02.015

Fig. 3.

Fig. 3

Phophoinositides inhbit the cleavage of WT ACTN4 by m-calpain between Y13 and G14. Phosphoinositides PI(4,5)P2, PI(3,4,5)P3, or PI(3,4) (50 µM each) were incubated with ACTN4 at room temperature for 30 min prior to the incubation with indicated amount of m-calpain for additional 1h at 30°C. Reaction was stopped by the addition of 1/5 reaction volume of 5× SDS sample buffer followed by 3 min boiling to denature proteins. Protein bands were separated by SDS-PAGE followed by either staining with Coomassie or immunoblotting of ACTN4 antibody aa 2–11. Graph represents the density of each band of aa 2–11 immunoblotting. Image represents one of three independent experiments.