Screening and identification of stable RORβ-overexpression GC cells

The RORβ gene coding sequence from RORβ/pReceiver plasmid (GeneCopoeia, Inc.) was inserted into pEGFP-C1 vector at the BamHI site and identified by DNA sequencing (Sangon Biotech Co., Ltd.). GC cells were seeded into 6-well plates at a density of 1×106 cells/well at 37°C with 5% CO2 for 24 h. RORβ/pEGFP-C1 (4 µg/µl) and pEGFP-C1 vector (4 µg/µl) were transfected into GC cells using Lipofectamine® 3000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.). After 2 days, the cells were cultured in the presence of 500 µg/ml G418 (Invitrogen; Thermo Fisher Scientific, Inc.) for 2 weeks to obtain stably transfected GC cells. RORβ-overexpression GC cells were further verified by western blotting.

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