Extended Data Fig. 4: Cryopreservation is ligand independent.
From: Mineralocorticoid receptor phase separation modulates cardiac preservation
Adeno-associated virus 9 (AAV) was administered to Myh6CreERT2;MRfl/fl mice 28 days prior to cardiac preservation. AAV9 was used to deliver cardiac-specific constructs containing MR with a deleted ligand binding domain (LBD, MRΔLBD). a, After 16 h of preservation followed by ex-vivo perfusion, construct expression in-vivo was verified by western blot. b, Murine donor hearts from wild type control, Myh6CreERT2;MRfl/fl mice infected with AAV-Luciferase (AAV-Luc) control, and Myh6CreERT2;MRfl/fl mice infected with AAV-MRΔLBD (MR missing the LBD) were cold (4 °C) preserved for 16 h followed by ex-vivo perfusion with Kreb buffer (n = 8). c, Immunostaining of MR condensates (red) with DAPI (blue). Representative fluorescence images of n = 4/group. d, Transcript expression of MR target genes ZFP219, CAMK2D, and PER1 in ex-vivo perfused murine hearts cold preserved for 16 h (n = 4 per group). Murine donor hearts were cold preservation with HTK only versus HTK + aldosterone (50 μM) (n = 8/group) for 12 h followed by ex-vivo perfusion. The cardiac functions e, contractility and f, relaxation were assessed by conduction catheter. Data are presented as means ± SD. *P < 0.05 and **P < 0.01 by two-sided Mann-Whitney test. Tukey’s corrections were used for multiple comparisons using one-way ANOVA for multiple group comparisons.
