Goat anti mouse a488

For immunostaining, mice were deeply anesthetized with Fatal-Plus and perfused with cold PBS, followed by 4% paraformaldehyde in PBS, pH 7.4. Brains were removed and post-fixed overnight at 4°C and submerged in 30% sucrose. Forty-micrometer-thick sections were cut on a sliding microtome, blocked in 5% normal goat serum and incubated overnight at 4°C in biotin-conjugated WFA (1:1000; Sigma-Aldrich L1516) to label PNNs, rabbit anti-PCP4 (1:500, SCBT, sc-74186) or mouse anti-regulator of G-protein signaling 14 (RGS-14) (1:500, UC Davis/NIH NeuroMab Facility, AB_10698026) to label CA2 pyramidal cells, or guinea pig anti-ZNT3 antibody (1:500, Synaptic systems #197 004) to label axons from the dentate gyrus (mossy fibers). Sections were washed three times in PBS and incubated in combinations of the following secondary antibodies at 1:500 for 40 min at room temperature: streptavidin Alexa-488 (Invitrogen #S11226), goat anti-rabbit A633 (Invitrogen #A21071), goat anti-mouse A488 (Invitrogen # A11008), or goat anti-guinea pig A633 (Invitrogen #A21105). Sections were mounted with Vectashield antifade mounting medium with DAPI (Vector Laboratories #H-1500). Images were acquired on a Zeiss laser scanning confocal (LSM510 NLO) or a Zeiss light microscope using controlled camera settings. Images from the different wavelengths were pseudo-colored for figure clarity.