RNA was extracted using the Quick-RNA MiniPrep Kit (Zymo, R2060) according to the instructions of the manufacturer including an additional DNase digestion. RNA concentrations were quantified via Qubit and cDNA was generated using the SuperScript IV First Strand Kit (Life Technologies, 18091200) according to the manufacturer’s instructions. SYBR green qPCR reactions were carried out in duplicates using POWERUP SYBR Green Master Mix (Life Technologies, A25743) and custom primer pairs (Supplementary Table 14) according to the manufacturer’s recommendations. Data were analyzed using the ΔΔCt method88 and GAPDH as reference. Between group differences were calculated using one-way ANCOVA with sex as covariate. Significance threshold was set at P = 0.05.
