DDX50 shRNA knock down or control shRNA in primary neonatal keratinocytes under differentiation conditions to determine the function of DDX50 in skin homeostasis.
Primary human keratinocytes were isolated from fresh surgically discarded newborn foreskin, and progenitor cells cultured in complete Keratinocyte-SFM (Life Technologies, #17005-142) and Medium 154 (Life Technologies #M-154-500). Cells were induced to differentiate by the addition of 1.2 mM calcium for 3 days at full confluence. Biological replicates were performed in all cases using primary cells from at least 2 independent, unrelated donors. Total RNA was extracted using the RNeasy Plus kit (QIAGEN, 74136), followed by poly(A) enrichment and library preparation using the NEB Ultra II (non-directional) kit (E6111).
NIAMS-phs003766
a_shDDX50-4_biorep3_kc211226