Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression
Figure Lengend Snippet: Generation of TWIK-1 BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.
Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).
Techniques: BAC Assay, Mouse Assay, Modification, Plasmid Preparation, Sequencing, Polymerase Chain Reaction, Slice Preparation, Expressing, Immunofluorescence