Biomechanical signals and the C-type natriuretic peptide counteract catabolic activities induced by IL-1β in chondrocyte/agarose constructs

Table 1 Description of the LNA probe and primer sequences used to quantify gene expression

Gene	Gene ID	Sequences	Product length	Efficiency
iNOS	4843	Probe: 5'-FAM-ACTTCTTTCCCGTCTCC-BHQ1-3'	305	1.99 ± 0.8
		Sense: 5'-TCCAGATAAGTGACATAAGTG-3'
		Antisense: 5'-CAGCTTGACCAGAGATTC-3'
COX-2	5743	Probe: 5'-AAACTGCTCAACACCG-BHQ1-3'	216	1.99 ± 2.8
		Sense: 5'- GGACAGGATTCTATGGAG-3'
		Antisense: 5'- GGATGTCAACACATAACTC-3'
Aggrecan	176	Probe: 5-'FAM-CCAACTCTTCAAGGTGA-BHQ1-3'	109	1.98 ± 0.4
		Sense: 5'-GACTGAAGTTCTTGGAGAA-3'
		Antisense: 5'-CACGAAAACCCAGAGTAA-3'
Collagen type II	1280	Probe: 5'-FAM-TCTGTCTCCTTGCTTGCCA-BHQ1-3'	200	1.99 ± 0.9
		Sense: 5'-GGAGTCAAGGGTGATCGT-3'
		Antisense: 5'-CTTGTGCACCAGCTTCTC-3'
GAPDH	2597	Probe: 5'-HEX-CAGTCAGCCGCATCTTCT-BHQ1-3'	160	1.98 ± 4.3
		Sense: 5'-TCTCTGCTCCTCCTGTTC-3'
		Antisense: 5'-CGCCCAATACGACCAAAT-3'

Primers used in PCR experiments with Locked Nucleic Acid (LNA) probes produced amplicons between 109 and 305 base pairs and efficiency values between 1.98 and 1.99. Probes contain fluorescein (FAM) or 6-carboxyhexafluorescein (HEX) as the 5'-reporter dye and Black Hole Quencher 1 (BHQ1) as the 3'-quencher. Nucleotides highlighted in bold denote the LNA base.

ISSN: 1478-6362