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Fig 1 Strategy and characterization of mouse QRFP gene disruption.

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posted on 2016-10-20, 01:16 authored by Kitaro OkamotoKitaro Okamoto, Miwako Yamasaki, Keizo Takao, Shingo Soya, Monica Iwasaki, Koh Sasaki, Kenta Magoori, Iori Sakakibara, Tsuyoshi MiyakawaTsuyoshi Miyakawa, Michihiro Mieda, Masahiko Watanabe, Juro Sakai, Masashi Yanagisawa, Takeshi SakuraiTakeshi Sakurai
A, Strategy for QRFP disruption. B, BamHI; E, EcoRI; H, HindIII; K, KpnI; S, SalI; X, XbaI; Xh, XhoI. GFP, green fluorescent protein; mPrm1, a part of second exon of the
murine protamine-1 gene, which contains an intron and a polyadenylation site
B, Immunohistochemistry (left panels) and in situ hybridization (right panels) of coronal sections of brains from wild type (upper panels) and QRFP-/- mice (lower panels),
showing complete depletion of QRFP expression in QRFP-/- mice. Insets in left panels show high power views of corresponding yellow rectangular region in each panel.
C, Distribution of QRFP neurons revealed by immunostaining with anti-GFP antibody in hypothalamic slice of Qrfp-/+ (GFP knock-in) mouse. C1, Coronal brain section of Qrfp-/+- mouse, stained with anti-GFP antiserum. C2, Higher power view of rectangular region shown in C1. C3, Higher power view of rectangular region shown in C2.
D, Immunostaining of hypothalamic slice of Qrfp-/+ brain with anti-GFP and anti-QRFP antibodies revealed that more than 95% of QRFP neurons were labeled with anti-GFP antibody, without any evidence of ectopic expression. D1, QRFP-like immunoreactivity. D2, Immunofluorescent image of GFP. D3, Merged image of D1 and D2

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