Tendon Gene Expression Analysis

Trizol/chloroform extraction was used to isolate RNA from dissected tendons. cDNA was then synthesized via reverse transcription using the SuperScript VILO master mix (Cat. # 11755050, Invitrogen, Carlsbad, CA). Gene expression was assessed by qRT-PCR using SYBR PCR Master Mix (Cat. # 4309155, Thermo Fisher, Waltham, MA) and calculated using the standard curve method or the 2^–∆∆Ct method relative to carrier-treated control tendons. The housekeeping gene, Gapdh, was used to normalize gene expression. Primer sequences for TGFβ-related molecules are listed in Supplementary file 1. All other primers were previously described (Howell et al., 2017 (link)).