hearts  (PromoCell)

Journal: Nature

Article Title: Cell-type-targeted mitochondrial transplantation rescues cell degeneration

doi: 10.1038/s41586-026-10391-0

Figure Lengend Snippet: a. Binder scaffolds used for mitochondria delivery. b. Schematic of full-length antibody conjugation to SNAP-tag-displaying mitochondria. c. HEK293T cells expressing a SNAP-tag at the outer membrane of mitochondria. The SNAP-tag is stained by anti-SNAP antibodies (magenta). Mitochondria are stained by anti-MT-CO1 antibodies (green). The SNAP-tag display on the mitochondrial surface was validated in at least three independent experiments. d. Immunostaining of isolated mitochondria displaying benzylguanine (BG)-conjugated anti-CD31 antibodies bound to the mitochondria outer membrane displayed SNAP-tag (bottom). Isolated mitochondria without BG-anti-CD31 antibodies are shown at the top. Mitochondria were detected by anti-SNAP-tag antibodies (magenta). Mitochondria-displayed anti-CD31 antibodies were detected with anti-mouse IgG conjugated with Alexa fluor-647 conjugated antibodies (cyan). e. Immunostaining of isolated mitochondria displaying BG-conjugated isotype control IgG1 bound to the mitochondria outer membrane-displayed SNAP-tag (bottom). Isolated mitochondria without BG-IgG1 (top). Mitochondria were detected by anti-SNAP-tag antibodies (magenta). Mitochondria-displayed IgG1 were detected with anti-mouseIgG conjugated with Alexa fluor-647 conjugated antibodies (cyan). f. Colocalization of SNAP-tag with either BG-anti-CD31 (left) or BG-IgG1 (right) in (d) and (e), respectively. n = 5, P < 0.0001, two-sided paired t test. g. Western blotting on isolated mitochondria displaying a SNAP-tag fused to a full-length antibody. Isolated mitochondria were incubated at different concentrations with either BG-conjugated anti-CD31 antibodies (BG-anti-CD31) or IgG1 (BG-IgG1). For controls, BG-free antibodies were incubated with isolated mitochondria displaying a SNAP-tag (SNAP-OMP25). Loading control, anti-TOMM20 antibody. A SNAP-tag interacting with the BG-conjugated antibody was detected with anti-SNAP-tag antibodies. Shifting in weight indicates SNAP-tag interaction with the light or heavy chain (LC/HC) of the BG-conjugated antibody. n × SNAP-OMP25, multiple SNAP-tag bound to LC or HC. For gel source data, see Supplementary Fig. . The experiment was repeated two times. h. Endothelial cells targeted by donor mitochondria displaying either IgG1 or anti-CD31 antibodies two hours after transplantation. Endothelial cells are stained by Phalloidin dye (green). Donor mitochondria displaying antibodies are stained by anti-SNAP antibodies (magenta). Cells are outlined with grey dashed lines. i. Quantification of the efficacy of the delivery of antibody-displaying mitochondria two hours after transplantation. n = 5, top: P = 0.1051 (10 nM), 0.0016 (100 nM), 0.0008 (500 nM) and bottom P = 0.7501 (10 nM), 0.0079 (100 nM), 0.0013 (500 nM), two-sided Welch’s t test and Mann-Whitney test (bottom 100 nM). Effect of avidity increase on mitochondria targeting efficiency (percentage) for anti-CD31: P = 0.0133 and for control IgG: P = 0.1432, Welch’s ANOVA test. Effect of avidity increase on mitochondria targeting efficiency (ratio) for control IgG: P = 0.0057, Welch’s ANOVA test. j. Schematic of mitochondria delivery displaying either BG-anti-CD31 or BG-IgG1 into primary endothelial cells (shown in magenta, CD31-positive) and cardiac fibroblasts (shown in brown, CD31-negative). k. Endothelial cells and cardiac fibroblasts targeted by donor mitochondria displaying either IgG1 or anti-CD31 antibodies two hours after transplantation. Endothelial cells are stained by anti-CD31 antibodies (red). All cells are stained with Phalloidin dye (green). Donor mitochondria displaying antibodies are stained by anti-SNAP antibodies (magenta). CD31-positive cells are outlined with grey dashed lines. l. Quantification of the efficacy of the delivery of antibody-displaying mitochondria two hours after transplantation. n = 4, P < 0.0001 (left) and P = 0.0158 (right), two-sided Welch’s t test. m. Immunostaining of tdTomato-expressing blood vessel organoids for CD31 (cyan), PDGFRβ (magenta), and RFP (red). The presence of vascular organoid cell types was validated in at least three induction batches. * P < 0.05, ** P < 0.01, *** P < 0.001. Data, mean ± s.e.m. Scale bars, 10 µm (c, d, e), 50 µm (h, k, m). The diagrams in b and j were created using BioRender; Ayupov, T. https://BioRender.com/fv9sxoi (2026).

Article Snippet: Human cardiac fibroblasts were obtained from LifeLine Cell Technology (FC-0060) and were maintained in FibroLife S2 Fibroblast Medium (Complete Kit, LifeLine Cell Technology, LL-0011) supplemented with 1% penicillin–streptomycin (Gibco, 15140-122).

Techniques: Conjugation Assay, Expressing, Membrane, Staining, Immunostaining, Isolation, Control, Western Blot, Incubation, Transplantation Assay, MANN-WHITNEY

Journal: PLOS One

Article Title: The novel mineralocorticoid receptor modulator balcinrenone protects against diet-induced cardiac microvascular dysfunction and plasma potassium elevation in mouse models

doi: 10.1371/journal.pone.0341078

Figure Lengend Snippet: Primary human cardiac fibroblasts were stimulated for 24 hours with or without 10 nM aldosterone, and balcinrenone or eplerenone added to final concentrations of 0.5, 2.5, or 12.5 µM. (A) Collagen 1 and (B) IL-6 concentrations in the supernatant were measured using an enzyme-linked immunosorbent assay. Data are presented as means ± standard deviations. * P < 0.05 versus control; # P < 0.05 versus 10 nM aldosterone. Aldo, aldosterone; Ctrl, control; IL-6, interleukin-6.

Article Snippet: Primary human cardiac fibroblasts (PromoCell, Heidelberg, Germany) were cultured in Fibroblast Growth Medium 3 (PromoCell, Heidelberg, Germany) according to the manufacturer’s instruction and used between passages five and seven.

Techniques: Enzyme-linked Immunosorbent Assay, Control