Quantifying Gene Expression in Femoral Head AC

Total RNA was isolated from femoral head AC using TRIzol reagent (Invitrogen, Waltham, Massachusetts). Complementary DNA (cDNA) was synthesized using a High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Waltham, Massachusetts). Quantitative real-time reverse transcription-polymerase chain reactions were performed in triplicate using a 7500 Real-Time qPCR system and SYBR Green reagents (Applied Biosystems, Waltham, Massachusetts). Specific primers used for qPCR gene expression analyses are listed in Table I (see supplementary Table i for a full list of gene names). Gapdh expression levels were used as internal controls. Gene expression levels were relatively quantified using 2-^∆∆Ct methods as described previously.^{7 (link),20 (link)}