WT and ΔN2-19 INX-6 containing GFP plus an 8×-histidine tag at the C terminus were subcloned into pGEM-HeFx and expressed in Xenopus oocytes by cRNA injection as described above. Uninjected oocytes were used for negative control. To examine whether ΔN2-19 blocks trafficking of WT, INX-6ΔN2-19 without tag was coexpressed with WT INX-6-GFP. Oocytes expressing INX6 proteins were washed with phosphate-buffered saline (PBS) and embedded with optimal cutting temperature (OCT) compound, and the frozen sections were prepared. Fluorescent micrographs were recorded using a BZ-X700 microscope (KEYENCE). To distinguish GFP signals at the plasma membrane from yolk autofluorescence, we used a color camera along with a GFP longpass filter.

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