mRNA expression in inflammed paws from wild type and GPR40 knockout mice

Bone marrow cells were isolated from femur and tibia from the hind legs of wild type and GPR40-knockout mice. The cells were passed through a cell strainer (70 µm), incubated in erythrocyte lysis (ACK) buffer for 5 minutes at room temperature, centrifuged, washed and resuspended in PBS-0,5% BSA. Unspecific binding was blocked by incubation with 60 µl of 2% Fc-blocking reagent Mouse BD Fc Bloc on ice. FACS sorting was performed using a FACS Diva (BD Biosciences) based on granularity and Ly6Ghigh expression (anti-Ly6G APC-cy7 (1A8)). RNA isolation, sequencing, quantification of mapped reads and differential transcript expression analysis was done by GenXPro GmbH (Frankfurt, Germany).