Quantification of CYP19A1 and ESR1 in Testes

Western blotting for CYP19A1 and ESR1 was performed as reported previously (45 (link)) on 80-d-old control and LCARKO testes. Blots were probed with primary antibodies ESR1 (sc-542; Santa Cruz Biotechnology, Dallas, TX, USA) and tyrosinated TUBA (α-tubulin) isoforms (ab6160; Abcam, Cambridge, MA, USA) or CYP19A1 [aromatase (Arom), ab18995; Abcam] and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (ab9486; Abcam). Primary antibodies were detected using either donkey anti-rabbit IRDye 680RD and goat anti-rat IRDye 800CW or donkey anti-goat IRDye 680RD and donkey anti-rabbit IRDye 800CW (all LI-COR Biosciences, Lincoln, NE, USA). Detection was carried out using the LI-COR Odyssey imaging system (LI-COR Biosciences) according to the manufacturer’s instructions.