Pik93

All salts were from Sigma-Aldrich. PAO was from Sigma-Aldrich, and wortmannin and PIK93 were from TOCRIS Bioscience. The following plasmids were used: GFP-PH-OSBP (Balla et al., 2005 (link); gift from Tamas Balla, National Institutes of Health, Bethesda, MD), GFP-PHx2-OSH2 (Stefan et al., 2011 (link); plasmid 36095; Addgene), GFP-P4M-SidM (Hammond et al., 2014 (link); plasmid 51472; Addgene), mRFP-PH-PLCδ1, mRFP-PH-Akt, GFP-Sac2 (Nakatsu et al., 2015 (link); gift from Pietro DeCamilli, Yale University, New Haven, CT), GFP-Rab27a, GFP-Rab3a, NPY-GFP, NPY-mCherry, VAMP2-pHluorin (all gifts from Sebastian Barg, Uppsala University, Uppsala, Sweden), Lamp1-RFP (Sherer et al., 2003 (link); plasmid 1817; Addgene), GFP-Rab5, GFP-EEA1 (gifts from Pietro DeCamilli), CIBN-CAAX (Idevall-Hagren et al., 2012 (link)), GFP-CRY2-5ptase_OCRL (Idevall-Hagren et al., 2012 (link)), GFP-CRY2-iSH2 (Idevall-Hagren et al., 2012 (link)), R-GECO1 (Zhao et al., 2011 (link); plasmid 32444; Addgene), GFP-Lact-C2 (Yeung et al., 2008 (link); plasmid 22853; Addgene), EGFP-Rab4a (Rzomp et al., 2003 (link); plasmid 49434; Addgene), GFP-Rab10 (Huckaba et al., 2011 (link); plasmid 31737; Addgene), Granuphilin-GFP (Gálvez-Santisteban et al., 2012 (link); plasmid 40032; Addgene), and GFP-2xFYVE_EEA1 (Petiot et al., 2003 (link)).

Phosphate buffered saline (PBS), medium-199, rat ELISA kits for GH, horse serum, penicillin, and streptomycin were purchased from Life Technologies (Grand Island, NY, USA). Fura 2-AM, the secondary Alexa Fluor 568 donkey anti-rabbit antibody, and 4′,6-diamidino-2-phenylindole (DAPI) were from Invitrogen (Eugene, OR). Rodent LH ELISA Kit was from Endocrine Technologies (Newark, CA). The primary antibody and standard for the PRL radioimmunoassay as well as the rabbit anti-PRL antibody for immunocytochemistry were from the National Pituitary Agency and Dr. A. F. Parlow (Harbor-UCLA Medical Center, Torrance, CA). ¹²⁵I-Prolactin was from Perkin Elmer Life Sciences (Boston, MA). Bovine serum albumin (BSA) fraction V and saponin were from MP Biomedicals (Solon, OH). The transcriptor first strand cDNA synthesis kit was from Roche Applied Sciences (Indianapolis, IN), Formaldehyde solution was from Thermo Fisher Scientific (Rockford, IL); Bay K8644, PIK93, UNC3230, and Wm were from Tocris (Bristol, UK). ISA2011B was from Med Chem Express (Monmouth Junction, NJ). TRH and dopamine were from Bachem Americas (Vista, CA). Unless stated otherwise, all other chemicals were obtained from Sigma (St. Louis, MO, USA).