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Electroporation
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Ldb1 and Rnf12-dependent regulation of Lhx2 controls the relative balance between neurogenesis and gliogenesis in retina
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Ldb1 and Rnf12-dependent regulation of Lhx2 controls the relative balance between neurogenesis and gliogenesis in retina
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Article Snippet
"Antibodies utilized for fluorescent immunohistochemistry are as follows: goat anti-Brn3 (1:200; Santa Cruz Biotechnology), mouse anti-calbindin (Calb1) (1:200; Sigma-Aldrich), rabbit anti-Calretinin (Calb2) (1:200; Chemicon), goat anti-Chat (1:100; Chemicon), sheep anti-Chx10 (Vsx2) (1:200; Exalpha Biologicals), rabbit anti-Dab1 (1:200; EMD Millipore), rabbit anti-DsRed (1:500; Clontech Laboratories), rabbit anti-GABA (1:200; Sigma), mouse anti-Gad6 (Gad2) (1:200; Developmental Studies Hybridoma Bank, University of Iowa), goat anti-GFP (1:500; Rockland Immunochemicals), rabbit anti-GFP (1:1000; Invitrogen), mouse anti-Glutamine synthase (Glul) (1:200; BD Biosciences), rat anti-Glycine (1:200; ImmunoSolution), mouse anti-Islet1 (1:200; Developmental Studies Hybridoma Bank), mouse anti-Ki67 (1:200; BD Biosciences), rabbit anti-Lhx2 (1:1500; generated in house with Covance), mouse anti-P27 (1:200; Invitrogen), mouse anti-Pax6 (1:200; Developmental Studies Hybridoma Bank), rabbit anti-TH (1:500; Pel Freez), mouse anti-VGlut3 (1:200; Antibodies Incorporated)."
Figure Legend
"Supplemental Figure 1. Electroporation of Lhx2 promotes the formation of wfACs. (a-c) Morphology of a wfAC generated following electroporation of Lhx2 . (d) Generated wide field amacrine cells co-express the pan-amacrine marker PAX6. (e-m) Co-labeling with amacrine cell subtype selective markers reveals that amacrine cells generated by Lhx2 electroporation do not fall within any well-established molecular category. ISLET 1, CHAT-cholinergic starburst amacrine cells; GABA, GAD2-GABAergic amacrine cells; GLYCINE-glycinergic amacrine cells; VGLUT3-glutamatergic amacrine cells; CALB2-mixed population primarily AII amacrine cells, A19 amacrine cells, and non-AII glycine immunoreactive amacrine cells; TH-dopaminergic wide field amacrine cells; DAB1-AII amacrine cells. GCL, ganglion cell layer; INL, inner nuclear layer; outer nuclear layer; s inner plexiform layer sublamina. Scale bars, 50 ¼m (all panels). "
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