Journal: PLoS ONE
Article Title: Annexin A6 regulates catabolic events in articular chondrocytes via the modulation of NF-κB and Wnt/ß-catenin signaling
Figure Lengend Snippet: mRNA levels for articular cartilage markers (aggrecan, type II collagen (α1(II)) and catabolic markers (Cox-2, IL-6, iNOS, MMP-13) in vehicle-treated, Wnt3a-treated, IL-1ß-treated, or combined Wnt3a- and IL-1ß-treated human articular chondrocytes overexpressing AnxA6. Human articular chondrocytes were transfected with empty expression vector (EV) or expression vector containing full-length AnxA6 ( AnxA6 ). Twelve hours after transfection cells were serum-starved for 24 h followed by treatment with IL-1ß, Wnt3a, IL-1ß, or combined IL-1ß and Wnt3a for 24h. Levels of mRNA were determined by real-time PCR using SYBR Green and normalized to the level of 18S RNA. The mRNA levels are expressed relative to the level of vehicle-treated cells transfected with empty expression vector, which was set as 1. Data were obtained from triplicate PCRs using RNA from 3 different cultures ( n = 3). Values are the mean ± SD. (**p
Article Snippet: PCRs were performed with a SYBR Green PCR Master Mix kit (Applied Biosystems), with 95°C for 10 min followed by 40 cycles at 95°C for 15 s and 60°C for 1 min, and 1 cycle at 95°C for 15 s and 60°C for 1 min.
Techniques: Transfection, Expressing, Plasmid Preparation, Real-time Polymerase Chain Reaction, SYBR Green Assay