Duplex quantitative real-time polymerase chain reaction (qRT-PCR) on total RNA from whole retinas was performed and relative normalized mRNA levels were calculated using the ΔΔCt method as described in [42 (link)]. Validated TaqMan™ assays utilizing FAM-labeled probes (Supplemental Table S1) were combined with an Actb- (β-actin)-specific assay utilizing a VIC-labeled probe (primer limited formulation, Thermo Fisher).
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