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. 2013 Jul 26;65(8):2113–2122. doi: 10.1002/art.37989

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Copyright © 2013 by the American College of Rheumatology

Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.

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Effect of receptor inhibition on inducible nitric oxide synthase (iNOS) expression. A, Chondrocytes were treated with 10 ng/ml interleukin-1β (IL-1β) along with 3 μM sphingosine 1-phosphate (S1P), 100 ng/ml pertussis toxin (PTX) (inhibitor of G_αi/S1P₁), 10 μM JTE-013 (inhibitor of S1P₂), or 5 μM suramin (inhibitor of S1P₃) for 3 hours. Expression of iNOS mRNA was quantified by real-time reverse transcription–polymerase chain reaction (RT-PCR). B, Chondrocytes were transfected with small interfering RNA (siRNA) against S1P₂ or a scrambled control (scr), and 24 hours after transfection, chondrocytes were treated with 10 ng/ml IL-1β and 3 μM S1P for 3 hours. Expression of iNOS mRNA was quantified by real-time RT-PCR. Results are the mean ± SEM arbitrary units, normalized to the values for the housekeeping genes, in 3 cartilage samples. ∗ = P < 0.05; ∗∗ = P < 0.01 versus IL-1β–treated vehicle control.