Effect of TAFA4 on Human macrophage M2c polarization

We aimed to decipher the effect of the neuropeptide TAFA4 on human M2c macrophage polarization. We used monocytes isolated from peripheral blood of three independant donors and generated different polarisations in the presence or absence of TAFA4. Monocytes were cultured with M-CSF for six days to induce their differentiation in macrophages (Mf). On day six, the cells were left untreated or were treated for two days with dexamethasone to induce their M2c polarization. After differentiation and polarization, 100,000 cells from each condition and donor were sorted to remove debris and dead cells, then used for the sequencing. Overall design: Total RNA was then extracted with the RNeasy Plus Micro Kit (Qiagen) and stored at -80°C until use. We used the Smart-seq v4 Ultra Low Input RNA Kit and the Nextera XT DNA Sample Preparation Kit for library construction. Sequencing was performed with HiSeq 4000 technology (Illumina).