|
Fig. S5
EPO supplement attenuate kidney structure and split diagram function alteration via a EPOR-dependent manner in zebrafish. A. Intracardiac injection of hEPO normalized enlarged glomerulus (white arrow head) and shortened pronephric neck (white asterixis) in EPO MO. In contrast, pronephros alterations in EPOR MO. could not be rescued by hEPO injections. Morpholino injected zebrafish embryos were injected at 24 hpf with 1 nl control or 4 IU/mL hEPO solution into the heart. Light microscopy images show increased hemoglobin concentrations in EPO MO. embryos injected with hEPO as compared to EPO MO. embryos injected with control solution only. In EPOR MO. embryos, hEPO could not increase hemoglobin concentrations. White scale bar in pronephros structure: 200 μm. Black scale bar in O-dianiside stain: 500μm. B. Quantification of data shown in A done in three independent experiments for each group. (n = 34–51 embryos per group). ns = not significant, **p ≤ 0.01, ***p < 0.001. C. Elevated loss of 70 kDa dextran–FITC fluorescence at 24 hpi and 48 hpi could be normalized in EPO morphants injected with hEPO (n = 42–50 embryos per group). Mean ± standard error. Significance was given for EPO MO. + KCL against Control MO. + KCL as ***p < 0.001, and EPO MO. + KCL against EPO MO + hEPO as #p < 0.05. D. Elevated loss of 70 kDa dextran–FITC fluorescence at 24 hpi and 48 hpi could not be rescued in EPOR MO zebrafish embryos injected with hEPO (n = 40–44 embryos per group). The same Control MO. + KCL and Control MO. + hEPO were applied. Significance was given for EPOR MO. + KCL against Control MO. + KCL as ***p < 0.001, and EPOR MO. + KCL against EPOR MO + hEPO as not significant (ns). All data were analyzed using the Student's t-test. Mean ± s.e.m.