Anti pgrmc1

Immunostaining of the OE was performed on 12 μm coronal cryosections of paraformaldehyde-fixed tissue from adult C57BL6 mice on superfrost slides (Thermo Scientific, Menzel Gläser). After blocking with 1% fish gelatin in phosphate-buffered saline containing 0.1% Triton X-100, sections were incubated with primary antibody (anti-Paqr8 (Abcam): 1:100, anti-Pacr9 (Abcam): 1:50, anti-GPR30 (Abcam): 1:100, anti-Pgrmc1 (Sigma-Aldrich): 1:100) and fluorescently-labeled secondary antibody (Invitrogen, anti rabbit Alexa 456 nm, 1:100) dilutions in blocking solution. Stained sections were mounted in ProLong Antifade Gold medium (Molecular Probes). All fluorescence images were collected on a confocal laser scanning microscope (LSM510 Meta, Zeiss, Oberkochen, Germany). For the anti-Pgrmc1 antibody, the staining was performed on tissue sections following a heat-induced epitope antigen retrieval using citrate buffer (10 mM citric acid, 0.05% Tween 20, pH 6.0). Control experiments completed without addition of the primary antibody revealed a low level of background staining.