Expression of the P2Y4 Nucleotide Receptor in the NSCs of the Hippocampus
(A) Total RNA was prepared from cultured NSCs derived from the hippocampus and was used for reverse transcription. As a negative control, samples without reverse transcriptase (RT) were also prepared. PCR was performed with the resulting cDNAs.
(B) Monolayer-cultured NSC clones c and d were fixed with 4% PFA and used for immunofluorescence to examine the expression of the indicated proteins. Scale bar indicates 25 μm.
(C) Monolayer-cultured clone d NSCs were fixed and used for immunofluorescence to examine expression of the indicated proteins. Scale bar indicates 10 μm.
(D) Cells co-expressing P2Y4 with the indicated proteins were counted. The graph shows the mean of three independent experiments and the error bars indicate SE.
(E) Coronal sections were prepared from frozen mouse brains and used for immunofluorescence with indicated antibodies. Arrowheads indicate P2Y4-positive cells. Scale bar indicates 7.5 μm.
(F) Cells co-expressing P2Y4 and indicated proteins in (E) were quantified. The graph shows the mean of results from three mice and the error bars indicate SE.