Fig. 3: Regulation of CD36 and arteriogenic gene expression via PKD-1 signaling in vascular endothelial cells.

a HMVECi-D cells were cultured in an endothelial medium (DMEM medium with 5% MVGS and 5% FBS). After starvation in serum-free DMEM medium for 6–8 h, the cells were treated with 10 µM of LPA, and/or 1 or 2 µM of CRT0066101 in serum-free DMEM medium for 24 h under 5% CO₂ and 37 °C. The total RNA was extracted and the mRNA levels of arterial gene ephrin B2 were detected by RT-qPCR. b Primary lung endothelial cells (ECs) were isolated from the control and EC-specific pkd-1 knockout mice. Passage 1 ECs were used for extraction and purification of total RNA. mRNA levels of pkd-1 were detected by RT-qPCR and relative expression was compared between the control and pkd-1-deficient ECs. c Increased expression of cd36 in pkd-1-deficient lung ECs. Lung ECs were isolated from the control and EC-specific pkd-1 knockout mice. Passage 1 lung ECs were used for extraction and purification of total RNA. mRNA levels were detected by qPCR and relative expression was compared between ECs from the control and EC-specific pkd-1 knockout mice. d Decreased expression of arteriogenic genes in pkd-1-deficient lung ECs. Primary lung ECs were isolated from the control and EC-specific pkd-1 knockout mice. Passage 1 ECs were used for extraction and purification of total RNA. mRNA levels were detected by RT-qPCR and relative expression was compared between the control and pkd-1-deficient ECs. e The cell lysate was extracted from passage 1 primary lung ECs that were isolated from EC-specific PKD-1 and control mice for detection of protein expression by Western blotting. Triplicate experiments were performed and levels of protein expression assessed by densitometry with Image J. ^*P < 0.05, ^**P < 0.01, or ^***P < 0.001 vs. control.