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. 2017 Sep 15;7:11710. doi: 10.1038/s41598-017-12188-8

Figure 6.

Figure 6

Synthesis of ion channel KvAP in CECF based reaction format. (A) Quantitative analysis of protein yield by scintillation measurements. Error bars represent the standard deviation of triplicate analysis. (B) Autoradiography and In-gel fluorescence of translation mixture, supernatant 1 and microsomal fraction of KvAP sample. Fluorescence samples were labeled by supplementation with BODIPY-TMR-Lys tRNAPhe to reaction mixture and integration of lysine-dye-conjugate into protein sequence. Fluorescence analysis of labeled KvAP was accomplished by using a Typhoon Trio+ Variable Mode Imager (GE Healthcare) (excitation nm, emission filter). (C) Voltage dependent recordings of KvAP currents at different voltages (Voltage scheme shown in blue lines). (D) Continuous single-channel records of KvAP currents at +100 mV on DOTAP bilayer and all point histogram of the current trace. (E) The current versus voltage (I-V) plots of the KvAP channel incorporated in DOTAP lipid bilayer. All recordings were done under symmetrical conditions; the medium contained 500 mM KCl, 10 mM HEPES, and pH 7.45.