WT and miR-146a-/- mice received 200μl intraperitoneal (i.p.) injections of Isotype control antibody (200μg/mouse /treatment) (Rat IgG1, κ; Biolegend, cat#400427) or IFN-γ blocking antibody (200μg/mouse/treatment) (R4.6A2, Biolegend, cat#505707) on day 0, 4, 8, and 12.
For translational in vivo experiments, WT mice injected with melanoma in the tail vein on day 0, received anti-PD-1 antibody, i.p. on days 1, 4 8, 16, and 22. Control mice were treated in parallel with an Armenian hamster isotype control purchased from BioXcell, (cat# BE0091). Oligonucleotides inhibiting miRNA-146a (ThermoFisher mirVana™ miRNA Inhibitor, Cat#: 4464088 ID: MH10722) or scramble controls (ThermoFisher mirVana™ miRNA Inhibitor, Negative Control #1, Cat#: 4464079) were administered via tail vein injections on days 5 and 9. In vivo-jetPEI® (Polyplus, cat# 201-50G), facilitated in vivo delivery of oligonucleotides across cell membranes, for 60μg of oligonucleotides injected in 200μl volume of 5% Glucose/in vivo-jet-PEI, per product protocol.
For translational in vivo experiments, WT mice injected with melanoma in the tail vein on day 0, received anti-PD-1 antibody, i.p. on days 1, 4 8, 16, and 22. Control mice were treated in parallel with an Armenian hamster isotype control purchased from BioXcell, (cat# BE0091). Oligonucleotides inhibiting miRNA-146a (ThermoFisher mirVana™ miRNA Inhibitor, Cat#: 4464088 ID: MH10722) or scramble controls (ThermoFisher mirVana™ miRNA Inhibitor, Negative Control #1, Cat#: 4464079) were administered via tail vein injections on days 5 and 9. In vivo-jetPEI® (Polyplus, cat# 201-50G), facilitated in vivo delivery of oligonucleotides across cell membranes, for 60μg of oligonucleotides injected in 200μl volume of 5% Glucose/in vivo-jet-PEI, per product protocol.