Fig. 2: WWP2 dysfunction regulates macrophage infiltration and expansion in the heart at 7 days post Ang II-infusion.

a Schematic representation of the experimental setup. Sorted live CD45+ cells from LV underwent scRNA-seq analysis, and seven clusters are represented in the UMAP space. WT, WWP2^wt/wt; Mut/Mut, WWP2^Mut/Mut. In each experiment, mice were treated with either saline (control) or Ang-II (500 ng/kg/min, 7 days). b Bar plots represent the proportions of macrophage clusters (Fig. 2a) in WT and Mut/Mut following Ang-II treatment. Two-proportion z test, ***P < 0.001 Mut/Mut vs WT. c Expression of Ly6c gene in cardiac macrophages represented in the UMAP space. d Percentage of Ly6c^high cells in WT and Mut/Mut cardiac monocyte compartments, in control or Ang-II-treated mice. n = 5–8 for each group. e Representative flow cytometry for CD11b⁺/F4/80⁺ gate in Ang-II-treated WT and Mut/Mut mice (left). Quantification of cardiac macrophages in Mut/Mut compared with WT mice following Ang-II treatment (right). n = 6–9 for each group. f Chemokine ligand-receptor pairs (Molecule 1–2 pairs) derived by CellPhoneDB analysis in cardiac macrophages, P < 0.01 (left). Mean expression of Molecule 1–2 pairs are reported as color intensity, and statistical significance (−log₁₀(p value)) as bubble size. Differential expression of chemokine ligands and receptors is represented as log₂fold changes (FC) (right). Two-sided Wilcoxon test. *, Benjamini–Hochberg adjusted P < 0.05. g–h Chemokine ligand (Ccl5, Ccl7, Ccl12, Ccl24) mRNA expression in macrophage clusters from scRNA-seq analysis, and in cardiac macrophages sorted from LVs of control or Ang-II-treated WT and Mut/Mut mice (n = 4–9 for each group). i–j Representative western blot (i) and quantitative RT-PCR (qRT-PCR) (j) measuring Ccl5 in bone marrow-derived macrophages (BMDMs) and spleen-derived macrophages (SPMs) under LPS (100 ng/ml) and IFNγ (10 ng/ml) treatment (4 hrs) in WT and Mut/Mut experimental groups. Non-parametric Mann–Whitney U test, n = 6 for each group. k Levels of Ccl5 in the supernatant from WT and Mut/Mut BMDMs treated with LPS (100 ng/ml) and IFNγ (10 ng/ml) (4 hrs). n = 5–6 for each group. Unless otherwise indicated, data are shown as dot-plots with mean ± SD, and statistical significance is assessed by the non-parametric Mann–Whitney U test.