Differentiation of MRC-iPS-25 cells to neural stem/progenitor cells. (A) Phase-contrast images of MRC-iPS-25 cells cultured on a feeder layer of MEFs (left) and NSPC/iPSCs (right). (B) Immunofluorescence analysis of Towne-infected MRC-iPS-25 cells cultured under feeder-free conditions at 2 dpi stained with primary antibodies for pluripotent markers (Nanog or Oct-4) and HCMV IE1/IE2 proteins. Antigen proteins were detected with Alexa Fluor 488-conjugated goat anti-mouse IgG or Alexa Fluor 594-conjugated goat anti-rabbit IgG antibody. Nuclei were stained with DAPI. (C) Immunofluorescence analysis of NSPC markers Nestin, Sox2, and Pax6 in NSPC/iPSCs. NSPC/iPSCs were fixed and reacted with anti-Nestin (green), anti-Sox2 (red), and anti-Pax6 (red) antibodies, followed by detection with secondary antibodies. Immunofluorescence signals were obtained using a fluorescence microscope IX71. Representative results from three independent experiments are shown.