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. Author manuscript; available in PMC: 2015 Jun 1.
Published in final edited form as: Nat Struct Mol Biol. 2014 Nov 10;21(12):1082–1090. doi: 10.1038/nsmb.2915

Figure 6.

Figure 6

PIP3 binding activity of tCNT-1a is stronger than that of AKT-1 by two orders of magnitude. (a) Immunoblotting of 5.1 μl of GST-tCNT-1a and 2.9 μl of AKT-1-His6 proteins synthesized and radiolabeled in rabbit reticulocyte lysate (*RRL) and 1 pmol of GST-tCNT-1a or AKT-1-His6 proteins purified from bacteria (Bact) using an anti-GST or an anti-His6 antibody (see METHODS). (b) Autoradiogram of 1 pmol of 35S-Met-labeled GST-tCNT-1a and AKT-1-His6 proteins made in RRL shows comparable radioactive signal intensity. (c) Lipid binding assays of GST-tCNT-1a or AKT-1-His6. Radiolabeled (*) GST-tCNT-1a or AKT-1-His6 with the indicated concentrations were incubated with lipid membrane strips as described in Fig. 5.