Fig. 4.
Oxygen–glucose deprivation (OGD)-induced κ-opioid receptor (KOR) trafficking shared similar pattern of KOR agonist-induced KOR internalization. Neuro2A cells stably transfected with mouse κ-opioid receptor–tdTomato fusion protein (N2A-mKOR-tdT) cells were untreated in control (A), incubated with 1 μM salvinorin A (SA), a selective KOR agonist, for 1 h (B), or treated with OGD for 1 h (C). Cells were fixed, mounted with 4',6-diamidino-2-phenylindole (DAPI), and observed by an inverted fluorescence microscope. SA induced KOR internalization, with a decrease of mKOR-tdT in the cell surface and an increase of crescent-shaped mKOR-tdT in the cytoplasm, which was similar to OGD-induced mKOR-tdT trafficking on the left. Merged images on the right show that both the internalized KORs by SA and OGD were clustered in the periphery of the nucleus. Examples of mKOR-tdT are indicated by the yellow arrows. Scale bar: 20 μm.

Oxygen–glucose deprivation (OGD)-induced κ-opioid receptor (KOR) trafficking shared similar pattern of KOR agonist-induced KOR internalization. Neuro2A cells stably transfected with mouse κ-opioid receptor–tdTomato fusion protein (N2A-mKOR-tdT) cells were untreated in control (A), incubated with 1 μM salvinorin A (SA), a selective KOR agonist, for 1 h (B), or treated with OGD for 1 h (C). Cells were fixed, mounted with 4',6-diamidino-2-phenylindole (DAPI), and observed by an inverted fluorescence microscope. SA induced KOR internalization, with a decrease of mKOR-tdT in the cell surface and an increase of crescent-shaped mKOR-tdT in the cytoplasm, which was similar to OGD-induced mKOR-tdT trafficking on the left. Merged images on the right show that both the internalized KORs by SA and OGD were clustered in the periphery of the nucleus. Examples of mKOR-tdT are indicated by the yellow arrows. Scale bar: 20 μm.

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