Immunocytochemistry of purified N27-A and unpurified N27 cells for dopamine neuron markers TH and DAT.

Abstract

<p>The N27 cells were cultured on 8-well chamber slides and immunostained for the dopamine neuron markers TH <b>(A-D)</b> and DAT <b>(E-H).</b> Other wells were double-stained for TH and the neuronal marker Tuj1 <b>(I-L).</b> To image every cell in each well, the nuclear marker DAPI was added to all wells. <b>(A-B):</b> The purified N27-A clone showed strong TH staining in all cells as demonstrated by dual-staining with TH and DAPI. <b>(C-D):</b> The unpurified N27 cell mixture revealed that only a small fraction of the DAPI-labeled cells were TH positive. <b>(E-F):</b> In the purified N27-A clone, all cells had moderate DAT staining as shown with DAT and DAPI double staining. <b>(G-H):</b> In the unpurified N27 cell mixture, very few cells were positive for DAT immunostaining. <b>(I-J)</b>: In the purified N27-A clone, all cells were double-positive for Tuj1 and TH. <b>(K-L)</b>: While there were few TH-positive cells in the unpurified N27 cell mixture, most cells were Tuj1 positive, demonstrating that the mixed cell population was neuronal. Bar, 50 μm for <b>A-L</b>.</p