Fig. 6: PB@PM comprehensively blocks the crosstalk between pyroptosis, apoptosis, and necroptosis in vivo.

A PCA of RNA-seq data (Sham: orange; MIRI: green; PB@PM: blue; n = 5 biologically independent replicates). B Volcano plot of differentially expressed genes (DEGs; PB@PM vs. MIRI: red/blue = up/downregulated). The Wald test of DESeq2 (DESeq(dds) default test = “Wald”) was used, which is a two-sided test. Multiple comparisons correction: the FDR (False Discovery Rate) correction of the BH (Benjamini-Hochberg) method was computed to obtain P adjustment (P.adj.). C Venn diagram of shared DEGs between Sham-MIRI and PB@PM-MIRI comparisons. D GSVA of pathway enrichment across groups. E KEGG pathway enrichment of downregulated genes in PB@PM vs. MIRI. A one-sided hypergeometric test was used. Multiple comparisons correction: the FDR correction for the BH method was calculated to obtain P.adj. F, G GSEA networks (F) and protein interaction networks (G) for pyroptosis, apoptosis, and necroptosis. H GSEA plots showing marked downward trends of pyroptosis, apoptosis, and necroptosis in the PB@PM group compared to the MIRI group. A permutation test was used, based on weighted Kolmogorov-Smirnov-like statistics. It is a two-sided test (tests whether the enrichment scores are significantly deviated from zero, which can be positive or negative, corresponding to up- or down-regulated enrichment, respectively). Multiple comparisons correction: The FDR correction for the BH method was calculated to obtain P.adj. I Pearson correlation between pyroptosis, apoptosis, and necroptosis in the Sham (C), MIRI (I), and PB@PM (T) groups. Using psych R package, method = “spearman” function, t-test, two-sided test, and no correction for multiple comparisons. J Western blot of PANoptosis-related proteins (AIM2, ZBP1, Pyrin, ASC, FADD, Bax, Bcl-2, phosphorylated/total RIPK1/RIPK3/MLKL, cleaved/total caspase-1/3/8 and GSDMD) in ischemic myocardium (day 3; n = 5 biologically independent replicates). Source data are provided as a Source Data file.