Extended Data Fig. 3: Characterization of Pik3ca mutant-specific BEC clusters.
From: Angiopoietin–TIE2 feedforward circuit promotes PIK3CA-driven venous malformations
(a) Unsupervised trajectory inference analysis of dermal BECs from control and Pik3ca mutant mice using SLINGSHOT, visualized in a UMAP plot. (b) Proportion of BECs from control and Pik3ca mutant mice expressing either the endogenous mouse Pik3ca transcript (wt) and/or the transgenic Pik3caH1047R transcript (H1047R). (c) Volcano plots showing results from DEG analysis between Pik3caH1047R transcript-positive and -negative BECs from the mutant mice within Pik3ca-1 and Pik3ca-2 clusters. Negative log2 fold changes (FC) represent upregulated gene expression in Pik3caH1047R transcript-negative BECs (wt /-), while positive log2 FC represent upregulated gene expression in transcript-positive BECs. Significance of differential gene expression was assessed using Seurat’s Wilcoxon rank-sum test. Note only a few genes significantly upregulated in the transcript-positive BECs, marked in red (P-value < 0.00001, log2FC ≥ 1). (d) ICAM1 expression in EMCN+ venous ECs in whole-mount ear skin from 4-OHT-treated Pik3caH1047R;Vegfr1-CreERT2 and Cre- littermate control mice (Ctrl). αSMA is used to label SMCs. Arrows indicate larger veins; arrowheads point to venous capillaries. Similar results were obtained from 3 mice in 1 experiment. (e) UMAP representation of Angpt2 transcript levels in BECs from control and Pik3ca mutant mouse. Scale bars: 50 μm.
