Transfection of lentiviral vectors with shRNA for GPR120 and GPR40

The pGMLV-GFP-vshRNA-GPR120 was constructed (pGMLV, a shRNAi Vector, Shanghai Genomeditech Co. Ltd, Shanghai, China). In the present study, we constructed 3 vshRNA-GPR120 lentiviral vectors (pGMLV-GFP-shRNA-GPR120) and 3 vshRNA-GPR40 lentiviral vectors (pGMLV-GFP-shRNA-GPR40) to silence the expression of GPR120 and GPR40 in murine BMMSCs (BMMSCs-vshRNA-GPR120/40). Three shRNA-targeting sequences for GPR120 were as follows: #1, 5′-GCACCCACTTCCCTTTCTTCT-3′ #2, 5′-GCTCTTCTACGTGATGACAAT-3′ and #3, 5′-GGACCAGGAAATTCCGATTTG-3′. And three shRNA-targeting sequences for GPR40 were as follows: #1, 5′-CCCTGCCCGACTCAGTTTC-3′ #2, 5′-GGCAGCCCACATAGCAGAA-3; and #3, 5′-CCGGGCCCGTCTCAGTTTCTCCATTCTCGAGAATGGAGAAACTGAGACGGGCTTTTT-3′. Stably transfectantclones were characterized by RT-PCR, western blot, and immunoblotting.