Fig 1: Decreased cardiomyocytes in pr130-/- zebrafish. (A) Representative images of H&E staining of adult heart tissue at 40× magnification. Scale bars, 200 µm; (B) Higher magnification of the boxed regions (400× magnification). The heart tissues of WT, M1, and M2 were from the same region of heart. Scale bars, 20 µm; (C) Pr130-/- exhibited reduced cardiomyocytes. Red arrows indicate the myocardium nucleus. Scale bars, 10 µm; (D) The cardiomyocytes in heart tissues of controls and pr130-/- adult zebrafish were quantified and presented in the bar graph. We calculated the number of cardiomyocytes in each of four chosen fields per fish at a magnification of 1000×. n = 16 hearts analyzed per panel. Data represent the mean ± SD. *, p < 0.05.
Fig 2: Expression patterns of pr130 in zebrafish. (A) Expression of pr130 at different stages in whole fish during zebrafish embryogenesis and in 72 hpf-heart; (B) Relative expression of pr130 in non-cardiac tissue and cardiac tissue of 72 hpf embryos was identified using real-time reverse transcription-polymerase chain reaction (RT-PCR); (C) The expression of pr130 in different tissues of adult zebrafish. Data represent the mean ± SD. **, p < 0.01.
Fig 3: Ultrastructure of adult heart tissues observed using transmission electron microscopy. Pr130-/- zebrafish displayed disordered myocardium. In pr130-/- heart, the boundary of bright band (red boxes), M line (yellow arrows), and Z line (red arrows) were not clear. In some cases the gaps between some longitudinal myocardial fibers increased (yellow boxes and white arrows).
Fig 4: Pr130 deletion causes defective function of heart. (A) The diastolic surface areas of ventricular and atrial; (B) Fractional shortening of ventricular; (C) Fractional area change of ventricular; (D) The systolic surface areas of atrial. V, ventricular; A, atrial. Data represent the mean ± SD. The symbols * and ** in the bar chart represent significant differences (p < 0.05 or p < 0.01). The numbers of zebrafish are indicated in the columns.
Fig 5: Effects of pr130 deletion on mortality and cardiac development. (A,B) pr130-/- embryos develop pericardial edema (arrow). A, WT; B, pr130-/-; (C) Survival curves; (D) Whole-mount in situ hybridization (WISH) using the heart probe cmlc2. Mutants showed a higher proportion of no-looping (arrows) compared with WT which showed normal heart looping; (E) Percentage of abnormal and normal heart looping. n = 100 embryos analyzed per panel. The symbols ** in the bar chart represent significant differences (p < 0.01).
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