Plasmid Preparation:Article Title: The PI(4)P phosphatase Sac2 controls insulin granule docking and release.
Article Snippet: PAO was from SigmaAldrich, and wortmannin and PIK93 were from TOCRIS Bioscience. .. The following plasmids were used: GFP-PH-OSBP (Balla et al., 2005; gift from Tamas Balla, National Institutes of Health, Bethesda, MD), GFP-PHx2-OSH2 (Stefan et al., 2011; plasmid 36095; Addgene), GFP-P4M-SidM (Hammond et al., 2014; plasmid 51472; Addgene), mRFP-PH-PLCδ1, mRFP-PH-Akt, GFP-Sac2 (Nakatsu et al., 2015; gift from Pietro DeCamilli, Yale University, New Haven, CT), GFP-Rab27a, GFP-Rab3a, NPY-GFP, NPYmCherry, VAMP2-pHluorin (all gifts from Sebastian Barg, Uppsala University, Uppsala, Sweden), Lamp1-RFP (Sherer et al., 2003; plasmid 1817; Addgene), GFP-Rab5, GFP-EEA1 (gifts from Pietro DeCamilli), CIBN-CAAX (Idevall-Hagren et al., 2012), GFP-CRY2-5ptaseOCRL (Idevall-Hagren et al., 2012), GFP-CRY2iSH2 (Idevall-Hagren et al., 2012), R-GECO1 (Zhao et al., 2011; Nguyen et al. Journal of Cell Biology 3724 Sac2 regulates insulin secretion https://doi.org/10.1083/jcb.201903121 D ow nloaded from http://rupress.org/jcb/article-pdf/218/11/3714/1608122/jcb_201903121.pdf by Philippines user on 24 February 2025 plasmid 32444; Addgene), GFP-Lact-C2 (Yeung et al., 2008; plasmid 22853; Addgene), EGFP-Rab4a (Rzomp et al., 2003; plasmid 49434; Addgene), GFP-Rab10 (Huckaba et al., 2011; plasmid 31737; Addgene), Granuphilin-GFP (Gálvez-Santisteban et al., 2012; plasmid 40032; Addgene), and GFP-2xFYVEEEA1 (Petiot et al., 2003). .. mCherry-Sac2 was generated by restriction enzyme digestion to release full-length mouse Sac2 from Sac2-GFP followed by ligation to mCherry-C1 (Clontech).
Article Title: The PI(4)P phosphatase Sac2 controls insulin granule docking and release
Article Snippet: PAO was from Sigma-Aldrich, and wortmannin and PIK93 were from TOCRIS Bioscience. .. The following plasmids were used: GFP-PH-OSBP ( ; gift from Tamas Balla, National Institutes of Health, Bethesda, MD), GFP-PHx2-OSH2 ( ; plasmid 36095; Addgene), GFP-P4M-SidM ( ; plasmid 51472; Addgene), mRFP-PH-PLCδ1, mRFP-PH-Akt, GFP-Sac2 ( ; 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 ( ; plasmid 1817; Addgene), GFP-Rab5, GFP-EEA1 (gifts from Pietro DeCamilli), CIBN-CAAX , GFP-CRY2-5ptase OCRL , GFP-CRY2-iSH2 , R-GECO1 ( ; plasmid 32444; Addgene), GFP-Lact-C2 ( ; plasmid 22853; Addgene), EGFP-Rab4a ( ; plasmid 49434; Addgene), GFP-Rab10 ( ; plasmid 31737; Addgene), Granuphilin-GFP ( ; plasmid 40032; Addgene), and GFP-2xFYVE EEA1 ( ). .. mCherry-Sac2 was generated by restriction enzyme digestion to release full-length mouse Sac2 from Sac2-GFP followed by ligation to mCherry-C1 (Clontech).
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