Gene expression profile of monocytes from healthy donor and triple negative breast cancer (TNBC) co-cultured with FAP and CD29 [bulk RNA-seq]

These samples were either treatment naive or samples co-cultured with FAP and CD29. Myeloid cell subpopulation comprises denditric cells (DC), monocytes and macrophages. We used RNA sequencing to analyze the response of monocytes to FAP/CD29 co-culture. Magnetically-enriched circulating CD14+ monocytes (CD14 Microbeads-Miltenyi) from HD (N=3) and TNBC (N=3) patients were co-culture with FAP+CD29+. After 3 hours of culture FACS-sorted monocytes were isolated by FACS-ARIA III (BD) cell sorter directly collected on lysing TCL buffer (Qiagen) containing 1%-mercatpoethanol before storage at -80°. RNA extraction was performed from ex vivo circulating isolated CD14+ and FAP+ CD29+ co-cultured monocytes by Single Cell RNA purification Kit (Norgen#, 51800) according to the manufacturer’s protocol