Gene expression profile in mouse articular cartilage under normoxia (21% O2) and hypoxia (3% O2), with or without TIMP-3
Gene expression profile in mouse articular cartilage under normoxia (21% O2) and hypoxia (3% O2), with or without TIMP-3
Cartilage from mouse hip joints (hip caps) was dissected and cultured (two caps per well in a 96-well plate with serum-free Opti-MEM) under standard conditions for 72 hours. Half the samples were transferred to hypoxia (3% O₂) for 24 hours, after which rhuTIMP-3 was added to subsets of both normoxic and hypoxic samples for 20 hours. Hypoxic samples were cultured for a total of 44 hours in hypoxia. Samples were homogenized in QIAzol with the Precellys lysing kit CK28R and frozen at -80. Total RNA was extracted with the miRNeasy kit (QIAGEN) with in-column DNA removal. The sequencing library was generated with the QuantSeq 3' mRNA-Seq Library Prep kit FWD for Illumina (Lexogen).