Wild-type, GLP1R-expressing, or GPR40-expressing HEK293 cells were seeded at a density of 30,000 cells per well in 96-well plates and treated with ASO or ASO conjugates as indicated for approximately 24 hours before harvest. Total RNA was extracted from cells or tissue samples using the RNeasy Micro Kit or Mini Kit (Qiagen). Complementary DNA (cDNA) was generated using a High-Capacity cDNA Reverse Transcription kit (Applied Biosystems). Reverse transcription qPCR analysis was performed with a QuantStudio 7 Flex using MALAT1 TaqMan assays (human Hs01054576 and mouse Mm01227912, Applied Biosystems) following the manufacturer’s instructions. Mouse FOXO1 was measured using the following primer probe set: forward sequence, CAAAGTCACATACGGCCAATCC; reverse sequence, CGTAACTTGATTTGCTGTCCTGAA; and probe, TGAGCCCTTTGCCCCAGATGCCTAT. The data were normalized against acidic ribosomal phosphoprotein P0 (36B4) expression measured using the following primer probe sets: HEK293 cells, CCATTCTATCATCAACGGGTACAA (forward sequence) and AGCAAGTGGGAAGGTGTAATCC (reverse sequence); mouse islets, GAGGAATCAGATGAGGATATGGGA (forward sequence), AAGCAGGCTGACTTGGTTGC (reverse sequence), and TCGGTCTCTTCGACTAATCCCGCCAA (probe sequence).

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