Most of these terms can typically be associated with the plasma membrane, including binding, signal transducer activity (Z > 14), and structural constituent of cytoskeleton (Z > 22). In the whole kidney, 7 terms were significantly enriched, including structural molecule (Z > 6) and transporter activity Dinaciclib (Z > 7) (Fig. 5b). In biological processes, 19 terms were significantly enriched in the
VEC plasma membrane fraction, including cell–cell adhesion (Z > 6) and protein transport (Z > 16). In the whole-kidney lysate, 5 terms were significantly enriched, such as metabolic process (Z > 28) and response to hormone stimulus (Z > 9) (Fig. 5c). In this study, we identified
16 PF299 concentration proteins known to be VEC marker membrane proteins in the CCSN-labeled plasma membrane fraction (Table 1). In addition, 8 proteins not previously reported to be VEC proteins in the kidney were confirmed to be VEC proteins on the basis of the immunolocalization of these orthologous proteins in the kidney as demonstrated in the Human Protein Crenigacestat Atlas (Table 2). Among these proteins, we focused on Deltex 3-like (Dll3), because growing evidence suggests that Dll families (Dll1, Dll3, and Dll4) act as Notch signaling ligands and participate in regulation of vasculogenesis and angiogenesis by modulating Notch signaling pathway . This has not been demonstrated previously in VECs of any Sclareol organ.
We then investigated the actual subcellular location of Dll3 by immunohistochemical and double-labeled immunofluorescence techniques using human kidney sections and anti-Dll3 antibody. The results of immunohistochemical analysis showed Dll3 expression in VECs specifically in kidney (Fig. 6a, b). Immunofluorescence microscopy showed co-localization of Dll3 and caveolin-1 to glomerular capillaries, veins, and arteries, but not to tubules elsewhere (Fig. 6c–k). Table 1 VEC membrane marker proteins identified in the VEC membrane fraction Prot Desc Accession No.a Prot_Matches Prot_Scoreb Prot_Sequence Mass cover (%) Dipeptidyl peptidase 4 IPI00208422 35 297 88,774 17.5 Carbonic anhydrase 3 IPI00230788 32 434 29,698 12.4 Sodium/potassium-transporting ATPase subunit alpha-1 IPI00326305 28 613 114,293 17.6 Integrin alpha-1 IPI00191681 16 149 91,687 12.9 Integrin beta-3 IPI00198695 11 118 90,066 10.9 Integrin beta-2 IPI00360541 9 113 87,955 13.7 Epidermal growth factor receptor IPI00212694 7 47 138,225 11.7 Scavenger receptor class B type 2 IPI00464469 7 56 56,705 7.3 Von Willebrand factor A domain-containing protein 5A IPI00400616 6 49 92,280 7.