Gp sirna mate plus transfection reagent

Chondrocytes were seeded on 6-well culture slides and grown to 70% confluence. The cells were transfected with 50 nM miR-100 mimic, 100 nM miR-100 inhibitor, or the corresponding control oligonucleotide (miR-100-mimic-NC or miR-100-inhibitor-NC) using the GP-siRNA-mate-plus transfection reagent (GenePharma, China) in 2 ml of α-MEM according to the manufacturer’s instructions. Six h post-incubation, the medium was removed, and cells were washed thrice with PBS and transfected for 48 h in standard conditions. The sequences were as follows: hsa-miR-100-5p mimic (sense, 5′-AACCCGUAGAUCCGAACUUGUG-3′; antisense, 5′-CACAAGUUCGGAUCUACGGGUU-3′), hsa-miR-100-5p inhibitor (sense, 5′-CACAAGUUCGGAUCUACGGGUU-3′), mimic negative control (sense, 5′-UUUGUACUACACAAAAGUACUG-3′; antisense, 5′-CAGUACUUUUGUGUAGUACAAA-3′), and inhibitor negative control (sense, 5′-CAGUACUUUUGUGUAGUACAAA-3′). All the oligos were synthesized by RiboBio (Guangzhou, China).

Chondrocytes were transiently transfected with siRNA targeting IL‐1R8 (GenePharma) or negative‐control siRNA (scrambled; GenePharma) using GP‐siRNA‐mate‐plus transfection reagent (GenePharma) following the manufacturer's instructions. All experiments were performed 48 hours after transfection, and the most effective single siRNA was used for further experiments.