Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Preclinical evaluation of cysteine protease-inhibitor aloxistatin (E64d) for heart failure therapy

doi: 10.1007/s00109-026-02695-5

Figure Lengend Snippet: Aloxistatin attenuates TGFβ1–induced fibroblast activation and ECM remodeling. A Heatmap of differentially expressed genes (DEGs; adjusted p -value ≤ 0.05, |log 2 fold change|≥ 1) between vehicle (veh) control and transforming growth factor β1 (TGFβ1)-treated HCFs with ICM and DCM backgrounds. B Overrepresentation analysis in DEGs in HCFs after TGFβ1 stimulation using GO, KEGG, Reactome, and WikiPathways databases as references. Selected fibrosis-associated terms are highlighted. C Schematic illustration of experimental design for investigating the effect of aloxistatin on TGFβ1-stimulated HCFs. HCFs were simultaneously stimulated with TGFβ1 and treated with aloxistatin. After 48 h, HCFs were collected for RNA isolation. D Overlap of deregulated genes in HCF after TGFβ1 stimulation and aloxistatin treatment. Values represent the number of genes in each category. E Fold changes in expression of overlapping genes from D in TGFβ1-stimulated HCFs (x-axis) and aloxistatin-treated TGFβ1-stimulated HCFs (y-axis) compared to respective controls. F Relative mRNA expression of fibrosis-associated markers α-smooth muscle actin 2 ( ACTA2 ) and connective tissue growth factor ( CTGF ) after TGFβ1 stimulation and aloxistatin treatment ( n = 3). G Top 30 terms overrepresented in opposite DEGs from E aloxistatin with GO, KEGG, Reactome, and WikiPathways reference databases. H Relative protein level of secreted fibronectin 1 (FN1) and matrix metalloproteinase 2 (MMP2) of TGFβ1-stimulated and aloxistatin treated HCFs. DMSO, dimethyl sulfoxide; ICM, ischemic cardiomyopathy; DCM, dilated cardiomyopathy

Article Snippet: Pro-fibrotic stimulation was performed with 5 ng/mL TGFβ1 (130–095-067, Miltenyi Biotec, Bergisch Gladbach, Germany) or respective vehicle (veh) control (4 mM HCl containing 0.1% bovine serum albumin; 9048–46-8, Serva, Heidelberg, Germany).

Techniques: Activation Assay, Control, Isolation, Expressing

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Preclinical evaluation of cysteine protease-inhibitor aloxistatin (E64d) for heart failure therapy

doi: 10.1007/s00109-026-02695-5

Figure Lengend Snippet: Aloxistatin inhibits inflammatory signaling and lowers oxidative stress. A Venn-diagram showing the overlap of regulated genes in RNA sequencing of rat living myocardial slices (LMS) treated with aloxistatin for 4 h and 24 h compared to respective dimethyl sulfoxide (DMSO) control. B Fold change of inflammation-related candidates significantly downregulated by aloxistatin treatment at both investigated timepoints. C NFκB signaling reported by luciferase activity in human embryonic kidney cells after 24 h treatment with aloxistatin [10 µM, 100 µM] ( n = 3). D Human leukocyte antigen-DR isotype (HLA-DR) expression in interferon γ (IFNγ) [5 µg/mL] stimulated or/and aloxistatin-treated induced pluripotent stem cell-derived macrophages 24 h after treatment ( n = 3). E Reactive oxygen species (ROS) levels assessed by monitoring fluorescence intensity in DCFDA-stained HCF over time after H 2 O 2 stimulation or/and aloxistatin treatment (top). Area under curve (AUC) analysis of ROS levels over time (bottom; n = 3). F Comparison of significant term enrichment (references: GO, KEGG, Reactome, WikiPathways) in overrepresentation analyses from differentially expressed genes in TGFβ1-stimulated HCFs after aloxistatin-treatment (x axis) and LMS cultivated for 24 h under aloxistatin exposure (y axis). RFU, relative fluorescence units

Article Snippet: Pro-fibrotic stimulation was performed with 5 ng/mL TGFβ1 (130–095-067, Miltenyi Biotec, Bergisch Gladbach, Germany) or respective vehicle (veh) control (4 mM HCl containing 0.1% bovine serum albumin; 9048–46-8, Serva, Heidelberg, Germany).

Techniques: RNA Sequencing, Control, Luciferase, Activity Assay, Expressing, Derivative Assay, Fluorescence, Staining, Comparison