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Rebecca McGirr, Leonardo Guizzetti, and Savita Dhanvantari

sorting of proglucagon into secretory granules are yet to be identified. Endocrine and neuroendocrine cells are characterised by the presence of a regulated secretory pathway, with specific intracellular compartments and associated proteins for the sorting

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SN Lee, E Prodhomme, and I Lindberg

Prohormone convertase 1 (PC1) is a serine proteinase responsible for the proteolytic processing of many precursor proteins within the regulated secretory pathway. The activity of PC1 is potentially regulated by two endogenous inhibitors, the PC1 propeptide and proSAAS. Here we have investigated the effect of proSAAS and propeptide-containing constructs on PC1 carboxy-terminal processing and activity. In AtT-20 cells, proSAAS expression inhibited both C-terminal PC1 processing and proopiomelanocortin (POMC) processing under pulse/chase conditions. SAAS CT peptide-propeptide chimeric constructs had no effect on the cleavage of PC1 and POMC under pulse/chase conditions. However, a construct containing the propeptide alone reduced C-terminal PC1 processing under pulse/chase conditions and also inhibited POMC processing. In contrast, experiments using HEK293 cells transiently expressing PC1 plus the respective constructs demonstrated significant inhibition of zymogen processing and decreased C-terminal processing of PC1 by the SAAS CT peptide portion of the chimera. Our results suggest that the PC1 propeptide expressed in trans is able to act as an endogenous inhibitor of PC1, but that SAAS CT peptide-containing/propeptide constructs cannot function as effective inhibitors of precursor maturation in the regulated pathway.

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Domenico Bosco, Dominique G Rouiller, and Philippe A Halban

fluorescence-activated sorting using a FACStar-Plus cell sorter (Becton–Dickinson, San Jose, CA, USA), as previously described ( van de Winkel & Pipeleers 1983 , Rouiller et al. 1990 ), resulting in a population comprising 95–98% (insulin-positive) β

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Caroline Alfaia, Vincent Robert, Kevin Poissenot, Yves Levern, Daniel Guillaume, Shel-Hwa Yeo, William H Colledge, and Isabelle Franceschini

population at E16.5. The discovery of higher levels of Esr1 in the ARC Kiss1 cells of female fetuses prompted us to compare the direct estradiol responsiveness of ARC Kiss1 cells in vitro specifically sorted from male and female fetuses

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Kotaro Horiguchi, Tom Kouki, Ken Fujiwara, Motoshi Kikuchi, and Takashi Yashiro

Anterior pituitary cells of male S100b-GFP rats were dispersed as described previously ( Horiguchi et al . 2008 ). Dispersed cells were separated into GFP-positive and GFP-negative cells by a cell sorter (MoFlo XDP: Beckman Coulter, Inc., Fullerton, CA

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Nathalie Hinfray, Rafael Henrique Nóbrega, Morgane Caulier, Damien Baudiffier, Emmanuelle Maillot-Maréchal, Edith Chadili, Olivier Palluel, Jean-Marc Porcher, Rüdiger Schulz, and François Brion

/total fish wet weight×100). Expression of cyp17a1 and cyp19a1 genes in sorted testicular cell fractions To generate data on the cellular localization of testicular cyp17a1 and cyp19a1 expression with an independent approach, we used testes from vasa

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Wei Zhang, Xin-Hong Wang, Si-Feng Chen, Guo-Ping Zhang, Ning Lu, Ren-Ming Hu, and Hui-Ming Jin

-ac-LDL) was from Molecular Probes, Inc. (Carlsbad, CA, USA). CD117 MicroBead Kit and magnetic activated cell-sorting system (MACS) were from Miltenyi Biotec (Bergisch Gladbach, Germany). Annexin-V/PI kit was from Bender MedSystems Inc. (Burlingame, CA, USA

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Hyunju Chung and Seungjoon Park

was directly proportional to the number of living cells. Fluorescence-activated cell sorting (FACS) analysis Cell cycle distribution was examined by FACS analysis. 1×10 6 cells were collected and fixed with 3.7% paraformaldehyde. After

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Kechun Tang, Teresa Pasqua, Angshuman Biswas, Sumana Mahata, Jennifer Tang, Alisa Tang, Gautam K Bandyopadhyay, Amiya P Sinha-Hikim, Nai-Wen Chi, Nicholas J G Webster, Angelo Corti, and Sushil K Mahata

2012 ). One of the major functions of CgA is to sort proteins to the regulated secretory pathway ( Chanat et al . 1991 , Taupenot et al . 2002 , Bartolomucci et al . 2011 ). Although the role of CgA in sorting or folding of membrane proteins is yet

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Susanne Granholm, Pernilla Lundberg, and Ulf H Lerner

). Fluorescence-activated cell sorting (FACS) Crude bone marrow and BMM cells, obtained (by culturing bone marrow cells for 6 days in the presence of M-CSF) as described above, were washed with PBS/3% FBS and stained with antibodies (0.4 μg/10 6 cells) against