Journal: Cell stem cell
Article Title: Deconstructing Olfactory Stem Cell Trajectories at Single Cell Resolution
doi: 10.1016/j.stem.2017.04.003
Figure Lengend Snippet: (A) Schematic of the olfactory epithelium describing the constituent cells: horizontal basal cell (HBC, green), globose basal cell (GBC, blue), sustentacular cell (Sus, pink), olfactory sensory neuron (OSN, purple), microvillous cell (MV, dark blue), Bowman’s gland (yellow). (B) Immunohistochemistry for the HBC lineage tracer YFP (green) and SOX2 (magenta) shows basal resting HBCs in the wild type (WT) background (left panel) and asynchronous differentiation following Trp63 conditional knockout (cKO) (center, right). (C) YFP(+) cells were collected by FACS at the indicated times following tamoxifen administration from mice carrying the Krt5-CreER; Rosa26eYFP transgenes and either the Trp63+/+ (WT) or Trp63lox/lox (cKO) alleles. (D) Sox2-eGFP(+)/ICAM1(−)/SCARB1(−)/F3(−) cells were collected by FACS; this enriched for the GBC, INP, and MV fates over Sus cells. (E) Data from both experimental designs were combined, filtered, normalized, clustered, and used in downstream analyses. – Scale bars, 50 microns. See Figure S1.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Armenian Hamster: phycoerythrin (PE)-conjugated ICAM1 BD Pharmingen Catalog No. 565615 Rabbit: phycoerythrin (PE)-conjugated SCARB1 Novus Biologicals Catalog No. NB400-104PE Rabbit: F3 Bioss Antibodies Catalog No. BS-4690R Chicken: GFP/YFP Abcam Catalog No. ab13970 Goat: SOX2 Santa Cruz Biotechnology Catalog No. SC17320 Donkey anti-Goat: Alexa-594 conjugated secondary antibody Thermo Fisher Scientific Catalog No. A-11058 Donkey anti-Chicken: Alexa-488 conjugated secondary antibody Jackson ImmunoResearch Catalog No. 703-545-155 Donkey anti-Rabbit: Alexa-555 conjugated secondary antibody Thermo Fisher Scientific Catalog No. A-31572 Donkey anti-Rabbit: Alexa-568 conjugated secondary antibody Thermo Fisher Scientific Catalog No. A-21207 Donkey anti-Mouse IgG: Alexa-568 conjugated secondary antibody Thermo Fisher Scientific Catalog No. A-10037 Donkey anti-Mouse IgG: Alexa-647 conjugated secondary antibody Thermo Fisher Scientific Catalog No. A-31571 Donkey anti-Goat: Alexa-647 conjugated secondary antibody Thermo Fisher Scientific Catalog No. A-21447 Goat anti-Chicken: Alexa-488 conjugated secondary antibody Thermo Fisher Scientific Catalog No. A-11039 Goat anti-Mouse IgG: Alexa-633 conjugated secondary antibody Thermo Fisher Scientific Catalog No. A-21050 Goat anti-Rabbit: Alexa-568 conjugated secondary antibody Thermo Fisher Scientific Catalog No. A-11011 Goat anti-Armenian Hamster: Dylight 594 conjugated secondary antibody Jackson ImmunoResearch Catalog No. 127-515-099 Mouse: P63 (4A4) BioCare Catalog No. CM163B Goat: NEUROD1 Santa Cruz Biotechnology Catalog No. SC1084 Mouse: NTUB/TuJ1 Neuromics Catalog No. MO-15013 Rabbit: SOX9 Milipore Catalog No. AB5535 Armenian Hamster: CD54/ICAM1 BD Pharmingen Catalog No. 550287 Alkaline phosphatase-conjugated anti-digoxygenin Sigma Catalog No. 11093274910 Alkaline phosphatase-conjugated anti-fluorescein Sigma Catalog No. 11426338910 Bacterial and Virus Strains None.
Techniques: Immunohistochemistry, Knock-Out