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. 2024 Apr 5;12:RP91729. doi: 10.7554/eLife.91729

Figure 3. Endothelial cell (EC)-activating perturbations modestly shift cells into the EC3 subtype.

(A) The proportion of cells in 7-day control and 7-day IL1B treatment are shown per human aortic endothelial cell (HAEC) donor and cluster on the top and for 7-day control and 7-day TGFB2 on the bottom. (B) The proportion of cells in 7-day siSCR control and 7-day siERG knockdown are shown per HAEC donor and cluster. EC1 was omitted in (A) due to lack of cells in both conditions.

Figure 3—source data 1. Annotated western blots for Figure 3—figure supplement 3B where the leftmost six wells are shown from left to right as (1) the protein ladder (labeled in kD), (2) the lipofectamine transfected control, (3) the scrambled siRNA control, (4, 5) two lanes using different siRNAs against the TCF4 gene (not relevant to these studies), and lastly, (6) siRNA against ERG.
The ERG blot (middle) and the H3 blots (bottom) originated from the same gel and membrane, which was cut at about 30 kD, and each piece was blotted for either anti-ERG antibody or anti-H3 antibody.
Figure 3—source data 2. Original western blots for ERG, annotated and uncropped.
Lanes are the same as described in Figure 3—source data 1.
Figure 3—source data 3. Original western blots for H3, annotated and uncropped.
Lanes are the same as described in Figure 3—source data 1.
Figure 3—source data 4. Original western blots for ERG, unannotated and uncropped.
Lanes are the same as described in Figure 3—source data 1.
Figure 3—source data 5. Original western blots for H3, unannotated and uncropped.
Lanes are the same as described in Figure 3—source data 1.

Figure 3.

Figure 3—figure supplement 1. Principal component analysis using RNA-seq data.

Figure 3—figure supplement 1.

(A) Principal component analysis (PCA) of EC1-4 snRNA-seq samples +/-siERG or control across donor replicates. (B) PCA of EC1-4 snRNA-seq samples +/-TGFB2 or control across donor replicates. (C) PCA of EC1-4 snRNA-seq samples +/-IL1B or control across donor replicates.
Figure 3—figure supplement 2. Receptor expression profiles across endothelial clusters.

Figure 3—figure supplement 2.

(A) Feature plots of expression of TGFB pathway receptors: TGFBR1, TGFBR2, and ACVRL1. (B) Feature plots of IL1B pathway receptors: IL1R1, IL1R2, IL1RL1, and IL1RL2.
Figure 3—figure supplement 3. Validation of ERG knockdown.

Figure 3—figure supplement 3.

(A) qPCR results for ERG knockdown across donors. (B) Western blot representing a typical knockdown of siERG with the siRNA pools used in this study. TCF4 samples are irrelevant for the purpose of this study.