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Binding capacity of 14–3–3 to TBC1D4 increased with insulin as shown in (a). Following immunoprecipitation of TBC1D4, 14–3–3 binding capacity to TBC1D4 was evaluated by far-western overlay (b) using digoxigenin-labelled 14–3–3 protein. **p < 0.01 for insulin effect, n = 12