Table 1.
Reference gene candidates evaluated by this study and their qPCR amplification efficiency.
Fig 1.
Stability ranking of 8 candidate reference genes by Bestkeeper.
Higher stability index is associated with higher stability. (A) Early, highly avascular time point, P14.5. (B) Late time point where neovascularization is peaking, P20. (C) Comprehensive ranking of reference genes across both time points of P14.5.
Fig 2.
Stability ranking of 8 candidate reference genes by Normfinder.
Lower stability index is associated with higher stability. (A) Early, highly avascular time point, P14.5. (B) Late time point where neovascularization is peaking, P20. (C) Comprehensive ranking of reference genes across both time points of P14.5.
Fig 3.
Stability ranking of 8 candidate reference genes by geNorm.
Lower stability index is associated with higher stability. (A) Early, highly avascular time point, P14.5. (B) Late time point where neovascularization is peaking, P20. (C) Comprehensive ranking of reference genes across both time points of P14.5.
Fig 4.
Stability ranking of 8 candidate reference genes by RefFinder.
Lower stability index is associated with higher stability. (A) Early, highly avascular time point, P14.5. (B) Late time point where neovascularization is peaking, P20. (C) Comprehensive ranking of reference genes across both time points of P14.5.
Table 2.
Summary of reference gene rankings by 3 primary algorithms and RefFinder, a summary tool, at each timepoint, P14.5, P20, and a comprehensive ranking of both timepoints.
Fig 5.
The percentage difference of expression level of Alox-15 in neonatal rat retinas normalized by different reference genes compared to TBP normalization at P14.5 by percentage (A), by fold change (C) and at P20 by percentage (B), and by fold change (D). Fold change was calculated by the 2−ΔΔCt method. All results are expressed as relative amounts to the RA group of the respective time point.
Fig 6.
OIR and PIA animal models and experimental groups.
In OIR groups, every 24 hours, the oxygen concentration was switched between 50% and 10% FiO2 until P14 when they were removed to room air (21% FiO2). Phlebotomy to induce PIA occurred from P3- P20 on hyperoxia days only, to prevent variation in oxygen concentrations during the highly sensitive hypoxia periods. Hematocrit levels of controls and PIA pups are shown. Bottom panel shows the experimental groups and timing of EPO administration.