Patch-clamp studies. Whole-cell currents were recorded as reported previously [11]. The COS-7 EGF receptor substrate eps15 acetyl were placed in a 2.5 ml recording chamber and superfused with normal Tyrode’s solution containing (in mM): NaCl 135, MgCl2 1.1, CaCl2 1.8, KCl 5.4, Hepes 10, glucose 10, pH 7.4 adjusted with NaOH at room temperature (21–23 °C). Voltage-clamp experiments were performed in the whole-cell configuration of the patch-clamp method by use of an Axopatch-200 patch-clamp amplifier (Axon Instruments, Foster City, CA), using fire-polished borosilicate glass pipettes with resistance of 2–4 MΩ when filled with the normal pipette solution containing (in mM): K-aspartate 100, KCl 30, MgCl2 1, Hepes 5, EGTA 5, Mg-ATP 5, Na2-creatine phosphate 5, pH adjusted to 7.2 with KOH [11] and [12].
Immunolocalization. The expression of Kvβ1.3 protein was examined using a polyclonal antibody (rabbit anti-human Kvβ1.3) raised against full-length Kvβ1.3. Cells were permeabilized and stained with Kvβ1.3 antibody and Texas red (TR)-conjugated anti-rabbit IgG as a secondary antibody and images were acquired using a confocal microscope [13].
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Conversely the concentrations of copper in the liver
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