<p><b>Copyright information:</b></p><p>Taken from "Inorganic pyrophosphate generation by transforming growth factor-beta-1 is mainly dependent on ANK induction by Ras/Raf-1/extracellular signal-regulated kinase pathways in chondrocytes"</p><p>http://arthritis-research.com/content/9/6/R122</p><p>Arthritis Research & Therapy 2007;9(6):R122-R122.</p><p>Published online 22 Nov 2007</p><p>PMCID:PMC2246241.</p><p></p> Phenotypic characterization of chondrocytes. Total RNA was extracted from untreated rat chondrocytes and subjected to real-time polymerase chain reaction (PCR) analysis. The relative abundance of gene mRNAs was normalized to that of S29 mRNA. Results are presented in histograms as mean percentages (± standard deviation [SD]) over S29 value. Effect of TGF-β1 on Ank, PC-1, and TNAP mRNA levels. Total RNA was extracted from rat chondrocytes exposed to 10 ng/mL of TGF-β1 from 1 to 48 hours and subjected to real-time PCR analysis. The relative abundance of gene mRNAs was normalized to that of S29 mRNA. Results are expressed as mean percentages (± SD) over control values. Statistically significant differences from the control are indicated as *< 0.05. Effect of TGF-β1 on ANK or PC-1 protein levels. Total proteins were extracted from rat chondrocytes exposed to 10 ng/mL of TGF-β1 from 6 to 48 hours and subjected to Western blotting using polyclonal anti-ANK and anti-PC-1 antibody. The protein band intensities were quantified by densitometry from enhanced chemiluminescence immunoblots. The relative abundance of these proteins was normalized to that of β-actin protein and expressed as induction folds over control value. N.D., not detected; TNAP, tissue-nonspecific alkaline phosphatase
