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. 2017 Oct 20;10:340. doi: 10.3389/fnmol.2017.00340

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Copyright © 2017 Ceolin, Bouquier, Vitre-Boubaker, Rialle, Severac, Valjent, Perroy and Puighermanal.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of Wfs1-CreERT2:RiboTag:Fmr1^−/Y mouse line. (A) Double immunofluorescence for Wfs1 (magenta) and HA (cyan) counterstained with DAPI (upper panels) and triple immunofluorescence for Iba1 (magenta), HA (cyan), and GFAP (yellow) (lower panels) in the dorsal hippocampus of Wfs1-CreERT2:RiboTag mice. Scale bars: 400 μm. (B) Double immunofluorescence for RGS14 (magenta) and HA (cyan). The lack of co-localization indicates that HA is preferentially expressed in CA1 but not CA2 pyramidal cells in Wfs1-CreERT2:RiboTag mice. (C) Triple immunofluorescence for HA (cyan), CaMKIIα (magenta), GAD67 (magenta), Iba1 (magenta), and GFAP (yellow) in the CA1 area of the dorsal hippocampus of Wfs1-CreERT2:RiboTag mice. Scale bars: 20 μm. For all immunofluorescence analyses, three slices per mouse were used (n = 3–4 mice/staining). (D) Drawing illustrating HA expression (cyan) among the distinct CA1 hippocampal cell types. (E) Validation by qRT-PCR (ΔΔCT) of the enrichment of pyramidal cell markers (Slc1a1 and Wfs1; cyan bars) and de-enrichment of GABAergic (Gad1, Slc32a1; magenta bars, Sst, Npy, Pvalb, Calb2, Kcnip1, and Grm1; orange bars) and mossy cell markers (Calb2; orange bars) as well as glial cells markers (Gfap, Cnp, and Aif1; gray bars) after HA-immunoprecipitation from hippocampi of Wfs1-CreERT2:RiboTag mice. Data are expressed as the fold change comparing the pellet fraction vs. the input containing the mRNAs from all cellular types (n = 6 mice/genotype). (F) Double immunofluorescence for HA (cyan) and FMRP (magenta) in the dorsal hippocampus of wild-type (left panels) and Fmr1^−/y (right panels) Wfs1-CreERT2:RiboTag mice (n = 3 mice). Scale bars: 400 μm. High magnification images in CA1 subfield correspond to areas delineated by the yellow stippled squares. Scale bars: 50 μm. Cx, cortex; DG, dentate gyrus; cc, corpus callosum; s.o., stratum oriens; s.p., stratum pyramidale; s.r., stratum radiatum; s.l.-m. stratum lacunosum-moleculare; s.m, stratum moleculare; s.gr, stratum granulosum; h, hilus. ^*p < 0.05; ^**p < 0.01; ^***p < 0.001.