All siRNAs were purchased from Dharmacon (Lafayette, CO, USA). Untreated cells and cells treated with non-targeting scrambled siRNA (Dharmacon, Catalog Item D-001810-10-20) were used as controls. Isoform‐specific siRNA was custom designed using Thermo Scientific siRNA Design Center (Thermo Scientific, Rockford, IL, USA). siRNA specific sequence was 5’-GAGGAAGACCUGAAGGUUCAGCAUA-3’ for PD-L1. For scrambled siRNA, we used the commercial ON-TARGETplus Non-targeting Control Pool (catalog number D-001810-10-20) comprised of the following siRNA sequences: 5’-UGGUUUACAUGUCGACUAA-3’, 5’-UGGUUUACAUGUUGUGUGA-3’, 5’-UGGUUUACAUGUUUUCUGA-3’ and 5’-UGGUUUACAUGUUUUCCUA-3’.
Cells were incubated for 48 h in RPMI 1640 medium containing siRNA-PD-L1 dextran NPs (concentration of siRNA: 100 pmol/mL, N/P = 15). Cells were treated with NPs for 48 h, because this incubation period resulted in the most effective downregulation of the target genes. All transfections were carried out based on established protocols (20 (link)).
Cells were incubated for 48 h in RPMI 1640 medium containing siRNA-PD-L1 dextran NPs (concentration of siRNA: 100 pmol/mL, N/P = 15). Cells were treated with NPs for 48 h, because this incubation period resulted in the most effective downregulation of the target genes. All transfections were carried out based on established protocols (20 (link)).
Free full text: Click here
