Competitive inhibition of Egr transcription factor-DNA binding attenuates TNFα decreases in cartilage matrix transcripts. Chondrocytes were transfected with 2 μM double-stranded, phosphorothiol-modified oligodeoxynucleotides containing Egr mutant or consensus DNA binding sequences and were treated with vehicle (black bars) or TNFα (grey bars) for 24 hours. Total RNA was collected and analysed by quantitative real-time PCR for (a) Agc1, (b) Hapln1 and (c) Col2a1, or 18S transcript levels. Data were analysed by the ΔΔCt method to acquire matrix gene transcript levels relative to 18S. Data from cells transfected with the Egr mutant or consensus oligodeoxynucleotides were normalized to vehicle-treated cultures and were log-transformed prior to analysis by paired and Student's t tests. *Significant difference (P < 0.05 by paired t test) in transcript levels compared with vehicle-treated cells transfected with the same oligodeoxynucleotide. #Significant difference in transcript levels between cultures transfected with mutant or consensus Egr binding sequences and treated with TNFα (P < 0.05 by Student's t test). Results are displayed as the mean ± standard error of five independent experiments.
Rockel et al. Arthritis Research & Therapy 2009 11:R8 doi:10.1186/ar2595