Next Generation Sequencing identifies lymph node lymphatic endothelial cell genes regulated by RANKL

Purpose: The objective was to identify genes regulted by RANK signalling in lymph node lymphatic endothelial cells Method: Lymphatic endothelial cells were FACS sorted from peripheral lymph nodes of mice deficient for stromal RANKL (Ccl19 Cre Rankl flox/flox) versus littermate controls (Rankl flox/flox) and processed for bulk RNA sequencing Results: Upregulated and downregulated genes were identified Lymphatic cell mRNA profiles of 8 week old littermate (Rankl flox/flox) and stromal RANKL-conditional knock out (Ccl19 Cre Rankl flox/flox) mice were generated by deep sequencing. The sequence reads that passed quality filters and key genes were validated by qRT-PCR using TaqMan and SYBR Green assays on lymphatic cells isolated from Prox-1 Cre Rank flox/flox mice.