Extended Data Fig. 1: Establishment of mice lacking endothelial SR-B1 or PDZK1.
From: SR-B1 drives endothelial cell LDL transcytosis via DOCK4 to promote atherosclerosis
a, j, Schematics of the gene-targeting strategies to generate floxed alleles of SR-B1 (a) and Pdzk1 (j) to create SR-B1fl/fl and Pdzk1fl/fl mice. Exon 2 of SR-B1 and exons 2 and 3 of Pdzk1were floxed. The recombined alleles following introduction of Cre recombinase are also shown. b, k, Using primers depicted in a and j, PCR-based genotyping was performed on aortas with or without intact endothelial cells (ECs). Aortas were obtained from floxed mice and mice with SR-B1 or PDZK1 deleted selectively from the endothelium (SR-B1∆EC or PDZK1∆EC). In genotyping to evaluate SR-B1 excision (b), additional samples were obtained from SR-B1fl/fl mice expressing Cre recombinase globally (gCre). In genotyping to evaluate Pdzk1 excision (k), lung samples were also studied. c, l, qRT–PCR analysis of expression of SR-B1 (c) or Pdzk1 (l) in primary aortic endothelial cells (c, n = 7 and 3 for SR-B1fl/fl and SR-B1∆EC, respectively; l, n = 6 per group) and bone marrow-derived myeloid lineage cells (c, n = 5 and 4 for SR-B1fl/fl and SR-B1∆EC, respectively; l, n = 5 per group)). d–f, SR-B1 protein abundance (d, e, n = 4) and Alk1 transcript levels (f, n = 4) were evaluated in aortic endothelial cells from SR-B1fl/fl and SR-B1ΔEC mice; summary data for SR-B1 protein are in e. The uncropped versions of all immunoblots shown are provided in Supplementary Fig. 1. CNX, calnexin. g, m, Immunoblotting of SR-B1 (g) or PDZK1 (m) protein abundance in liver. h, n, Plasma cholesterol levels. For SR-B1 studies, n = 4, 3, 9 and 10 mice for bars from left to right; for PDZK1 studies, n = 4, 3, 6 and 6 mice. i, o, Representative lipoprotein profiles. Data are mean ± s.e.m.; in c, e, i, P values calculated by two-sided Student’s t-test; in h, n, P values calculated by ANOVA with Dunnett’s post-hoc test.
