Extended Data Fig. 10: Combination treatment induces transcriptional and protein-level changes in TAC hearts without significantly affecting inflammation or immune signalling.
From: Selective inhibition of stromal mechanosensing suppresses cardiac fibrosis
(A) Left: Western blot of phosphorylated YAP (S127) and total YAP in whole heart lysates from the different drug treatment groups: Sham, Sham S + P, TAC 4 wk, TAC 6 wk, S + P, S, P (n = 5, 5, 10, 9, 9, 8, 9 hearts per lysate, respectively). Right: snRNA-seq violin plot displaying module scores for major YAP/TAZ target genes (Ccn2 (CTGF), Ankrd1, Amotl2, Birc5, and Igfbp2) specifically in CFs. (B) Violin plots showing module scores for common genes associated with oxygen consumption rate (OCR, left) and extracellular acidification rate (ECAR, right). (C) Normalized mass spectrometry intensity values for SORBS2 (ArgBP2) in two published datasets of mouse TAC hearts41,66: Kuzmanov et al and Rudebusch et al. (n = 5 and 4 tissue samples, respectively). (D) Violin plot of Sorbs2 expression in the five groups. Horizontal bars indicate mean. Cells with >0 Sorbs2 expression are shown. (E) Gene Ontology enrichment analysis for SAR + PFD versus TAC hearts. Fisher’s exact test (with Benjamini-Hochberg adjustment; one-sided). (F) Dot plot of major pro-inflammatory cytokine genes in the heart. (G-I) CellChat-based inference of cell-cell communication mechanisms, showing changes in modes of signalling from immune cells to CF populations (immune→CF) in the SAR + PFD group relative to TAC (H,I). A permutation test (randomization; one-sided) was used to compute communication probability strength of ligand-receptor pairs. (J-K) Immunoblots and corresponding quantitation of SRC phosphorylation and key fibrosis-associated proteins αSMA and POSTN for each group: Sham, Sham S + P, TAC (4 wk), TAC (6 wk), S + P, S, P (n = 5, 5, 10, 9, 9, 8, 9 hearts per lysate, respectively). Immunoblots in panels A and K are representative of n = 3 experiments performed independently with similar results. Images are from two blots from the same samples from the same experiment, processed under identical conditions. For panel K, data from an additional experiment was normalized and pooled. P-values were calculated based on ordinary one-way ANOVA with Dunnett’s test (*’s) versus Sham; and the two-sided Student’s t-test (#’s). Data are mean ± SEM.
