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In response to a glucose bolus, a 2-fold increase in blood glucose levels was rapidly observed in WT and SKO mice treated with vehicle, with minimal fluctuation in plasma insulin levels ( Figures 4F, G

Constant insulin stimulation may strain the pancreas

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The resulting lysates were incubated with either magnetic beads conjugated to anti-Flag antibodies or protein A/G agarose beads coupled to the corresponding antibodies (Thermo Fisher Scientific, 88802) at 4 C for 3 h