Journal: bioRxiv

Article Title: Fasting reverses PLN R14del-mediated cardiomyopathy through lysosomal reactivation

doi: 10.64898/2026.03.24.713684

Figure Lengend Snippet: A. Volcano plot showing all significantly (p-value < 0.05) downregulated (n = 201) and upregulated (n = 433) genes in wild type (WT) and plna R14del zebrafish hearts with a log₂(fold change) higher than +/− 0.7. Each dot represents one gene; blue corresponds to downregulated genes and green to upregulated genes. The p-value cut off corresponds to 1.30 on the -log 10 axis. B. Fold enrichment of downregulated and upregulated GO terms identified using the DAVID online GO analysis tool, when comparing WT and plna R14del zebrafish hearts in a pseudo-bulk dataset. C. Representative images of immunofluorescent staining against DAPI, Pln and Lamp1 in one year old WT and plna R14del zebrafish. Data shown are representative for n = 8 WT and n = 8 plna R14del zebrafish hearts. Scale bar = 5 μm. White arrowheads indicate Pln and Lamp1 co-localization. D. Quantification of number of Lamp1 foci divided by cardiomyocyte (CM) area. Statistics were performed by two-tailed unpaired t-test. Data obtained from n = 8 WT and n = 8 plna R14del zebrafish hearts. E. Quantification of Pln puncta co-localized with Lamp1 foci. Data obtained from n = 8 WT and n = 8 plna R14del zebrafish hearts. F. Representative image of immunofluorescent staining against DAPI, PLN and LAMP1 in left ventricle (LV) from patients with end-stage cardiomyopathy carrying the PLN R14del variant. Dashed box high lights the zoomed in area. Data shown are representative for n = 4 individuals. Scale bars = 10 μm. White arrowheads indicate PLN and LAMP1 co-localization. G. Representative images of immunoelectron microscopy against Lamp1 (10 nm gold) in one year old WT and plna R14del zebrafish hearts. * Indicate endo-lysosomal compartments. Data shown are representative for n = 3 individuals. Scale bar = 200 nm. H. Representative images of DQ red BSA BSA ex vivo assay in one year old WT and plna R14del hearts. Dots represent functional lysosomes. Data shown are representative for n = 4 WT and n = 3 plna R14del zebrafish hearts. Scale bar = 10 μm. I. Quantification of number of DQ red BSA foci per 5000 μm 2 . Data points represent an individual measurement per image; multiple points originate from one individual. Data obtained from n = 4 WT and n = 3 plna R14del zebrafish hearts. Error bars indicate mean ± s.d. Statistics were performed by two-tailed unpaired t-test. J. Representative images of DQ red BSA ex vivo assay in WT cardiac slices treated with medium or thapsigargin (100 μM). Dots represent functional lysosomes. Data shown are representative for n = 3 per group. Scale bar = 10 μm. K. Quantification of number of DQ red BSA foci per 5000 μm 2 . Data points represent an individual measurement per image; multiple points originate from one individual. Data obtained from n = 3 zebrafish hearts per group. Error bars indicate mean ± s.d. Statistics were performed by two-tailed unpaired t-test.

Article Snippet: Sections were then blocked for 5 min with 1% BSA-c/VG and incubated for 1 hour at room temperature with a primary antibody against Lamp1 (1:10, Boster Biological Technology, DZ01398-1).

Techniques: Staining, Two Tailed Test, Variant Assay, Immuno-Electron Microscopy, Ex Vivo, Functional Assay