Figure 4

Characterization of rEag1-I and rEag2-I chimeric channels. (A) Schematic representation of the construction of rEag1-I and rEag2-I chimeras. For all schematic cartoons hereafter, rEag1 and rEag2 sequences are shown in red and black, respectively. (B) Representative K⁺ currents recorded from Xenopus oocytes over-expressing the indicated Eag constructs. Two-electrode voltage clamp parameters: the holding potential for rEag1 and rEag2 was -90 and -110 mV, respectively; the pulse protocol comprised 300-ms depolarizing test pulses ranging from -70 to +60 mV (rEag1) or from -100 to +40 mV (rEag2), with 10-mV increments. (C) Membrane localization of GFP-rEag1-I/rEag2-I channels in HEK293T cells. Scale bar, 10 μm. (D) Expression of GFP-rEag1-I/rEag2-I channels in DIV12 hippocampal neurons. Scale bar, 25 μm. (E) Quantification of the number of GFP puncta per neuron for GFP-rEag1, GFP-rEag1-I, GFP-rEag2-I, and GFP-rEag2. The number in parenthesis denotes the amount of neurons analyzed. (*: significantly different from GFP-rEag1; t-test, p < 0.05)(#: significantly different from GFP-rEag2; t-test, p < 0.05)