Transfection of ADAMTS4 siRNA to osteoarthritic chondrocytes and cartilage explants: Transfection of siRNA for ADAMTS4 and non-silencing oligonucleotide to osteoarthritic chondrocytes

Supplementary material 1 Real-time quantitative PCR: Sequences of the primers and TaqMan probe for ADAMTS1, 4, 5, 8, 9 and 15 were designed using Primer Express software (Applied Biosystems, Foster City, CA): forward primer 5’-CCATCCCAAGAGTATCACATGTCT-3’, reverse primer 5’-CACTATGACACAGCAATTCTTTTCAC-3’ and TaqMan probe 5’-FAM-CCCACACAAGTCCTGTC-MGB-3 for ADAMTS1; forward primer 5’-TCACTGACTTCCTGGACAATGG-3’, reverse primer 5’-ACTGGCGGTCAGCATCATAGT-3’ and TaqMan probe 5’-FAM-CATCTGCCTGTGACTTTCCCTG-TAMRA-3’ for ADAMTS4; forward primer 5’-CACTGTGGCTCACGAAATCG-3’, reverse primer 5’-GGAACCAAAGGTCTCTTCACAGA-3’ and TaqMan probe 5-FAM-CTTGGCCTCTCCCATGACGATTCCA-TAMRA-3’ for ADAMTS5; forward primer 5’-GCAGAACCACATCCTGACGTTAA-3’, reverse primer 5’-ACGATCAGCACTTTTACCACCAT-3’ and TaqMan probe 5’-FAM-CTACAAGCACCCCAGCATCAAGAATTCCAT-TAMRA-3 for ADAMTS8; forward primer 5’-TCCGAGACTGCCGTAGAAAGA-3’, reverse primer CCGACAAAACCTGAAGCAAAA-3’ and TaqMan probe 5’-FAM-TTAACTCAACAGATCGCATTGAGCAA-TAMRA-3’ for ADAMTS9; forward primer 5’-GGTGAAATACCGATCCTGCCA-3’, reverse primer 5’-GTAGCCGTTGAAAGCCTCACA-3’ and TaqMan probe 5’-FAM-TCAGCCTCCGGAAAGAGCTTCCG-TAMRA-3’ for ADAMTS15. The total gene specificity of nucleotide sequences chosen for the primers and probes, and the absence of DNA polymorphisms were ascertained by BLASTN and Entrez on an NIH web sites (http://www.ncbi.nlm.nih.gov/).