PIEZO1 activation and signalling is associated with inflammation permissive phenocopy of synovial resident macrophages in early arthritis
PIEZO1 activation and signalling is associated with inflammation permissive phenocopy of synovial resident macrophages in early arthritis
On the days of the extraction, the cells were washed with D-PBS in the 24 well plate. Then the cells were lysed using RLT buffer + 1% b-mercaptoethanol, at RT, mixing well.  PIEZO1 activation using Yoda1  Macrophages were differentiated from CD14 monocytes (from 6 healthy donors) as described for 6 days in complete RPMI supplemented with 20 ng/mL M-CSF, changing medium on day 3. On day 5 of differentiation, the cells were stimulated with 20 uM Yoda1, for 12 hours (overnight), before being lysed for RNA extraction.  RNA was extracted using RNeasy Microkit, following manufacture's instructions (Qiagen, 74004). Libraries were prepared by University of Glasgow, MVLS Shared Research Facility, for Illumina sequencing.