Human GLP1R or GPR40 was stably overexpressed in HEK293 cells (GLP1R-HEK293 and GPR40-HEK293). Wild-type HEK293 cells (American Type Culture Collection) were routinely cultured in Dulbecco’s modified eagle medium (DMEM; Gibco, #31966) with 10% fetal bovine serum (FBS; Gibco, #10270) and GLP1R-HEK293 and GPR40-HEK293 in the same medium supplemented with hygromycin B (100 and 200 μg/ml; Invitrogen, #10687). For gene expression analysis, cells were cultured in poly-d-lysine (PDL)–coated 96-well plates (Corning) at 37°C, 95% humidity, and 5% CO2 and then incubated with increasing concentrations of ASO or ASO conjugates for 24 hours, followed by RNA extraction and expression analysis. Cells for β-arrestin and receptor internalization assays were cultured according to the manufacturer’s protocol (DiscoverX Corporation, Fremont, CA).

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