RNA-seq profiling of mouse lung endothelial cells to investigate ENG-dependent and ENG-independent BMP9-induced target genes.
The aim of this experiment is to dissect endoglin (ENG)-dependent and ENG-independent BMP9 target genes. We have shown previously (PMID: 31431534) that BMP9 in complex with soluble endoglin (sENG:BMP9) has compromised signalling in Eng knockout (KO) mouse lung endothelial cells (MLECs), but not in wild type (WT) MLECs, suggesting sENG:BMP9 signalling in WT MLECs is mostly ENG-dependent. Hence we designed this experiment to compare  sENG:BMP9 signalliing in WT cells with prodomain bound BMP9 (pro:BMP9) signalling in Eng-KO cells to tease out ENG-dependent and ENG-independent BMP9 target genes. Overall design: MLECs were isolated from four independent Eng fl/fl:Rosa26-CRE:Immorto mice. Eng KO MLECs were generted by tamoxifen induction in vitro, along with vehicle treated cells to obtain wild type (WT) control. Treatment in WT cells include PBS-treatment and sENG:BMP9 treatment. Treatment in KO cells include PBS-treatment and pro:BMP9 treatment. All samples were processed in parallel for RNA extraction and RNAsequencing.