Zen lite 2011 microscope software

MODE-K cells were plated on collagen-coated coverslips 1 day prior to transfection. On day 2, cells were transfected using Lipofectamine (Invitrogen™, Life Technologies) according to the standard procedures and left to expand in a 37°C 5% CO₂ incubator for 24 h. On the day of staining, cells were incubated with 10% normal horse serum (NHS), and thereafter with a rabbit anti-HA antibody (Sigma-Aldrich) and a goat anti-rabbit Cy5 (Jackson ImmunoResearch). Cells were blocked with NHS, stained with PE-conjugated anti-FLAG (Prozyme), fixed in 4% paraformaldehyde, and stained with 4′,6-diamidino-2-phenylindole (DAPI). Cells were viewed using confocal microscopy (Zeiss LSM700 Inverted) and analyzed with ZEN lite 2011 microscope software (Carl Zeiss).

Methanol-Carnoy-fixed and paraffin embedded sections were stained with anti-Btnl1 rabbit polyclonal antiserum or pre-immune serum. Sections were incubated with TRITC-conjugated AffiniPure F(ab’)₂ fragment donkey anti-rabbit IgG (H+L) (Jackson ImmunoResearch, West Grove, PA). Sections were mounted in ProLong^® Gold antifade reagent containing 4′,6-diamidino-2-phenylindole (DAPI) (Molecular Probes^®, Life Technologies, Eugene, OR). Tissue sections were viewed using confocal microscopy (Zeiss LSM700 Inverted) and analyzed with ZEN lite 2011 microscope software (Carl Zeiss, Oberkochen, Germany).