Fig 2: Immunohistochemical localization of PAP in transverse sections of rat CP.Representative photomicrographs for double staining with PAP (green), and the type I cell marker NTPDase2 (A, red), the type II cell marker PLC-ß2 (B, red), or a-gustducin (C, red), or the type III cell marker SNAP25 (D, red), AADC (E, red), or 5HT (F, red) are shown for transverse sections. In the photomicrographs on the right, cryosections were treated with the first antibody-free solution. Immunoreactivity due to the second antibodies only was used as a negative control (N.C.). Arrowheads indicate the colocalization of PAP and taste cell markers. The nuclei were counterstained with Hoechst 33258 (blue). Scale bar, 50 µm.

Fig 3: Immunohistochemical localization of PAP in longitudinal sections of rat CP.Representative photomicrographs for double staining with PAP (green), and the type I cell marker NTPDase2 (A, red), the type II cell marker PLC-ß2 (B, red), or a-gustducin (C; red), or the type III cell marker SNAP25 (D, red), AADC (E, red), or 5HT (F, red) are shown for longitudinal sections. Arrowheads indicate the colocalization of PAP and taste cell markers. The nuclei were counterstained with Hoechst 33258 (blue). Scale bar, 50 µm.

Fig 4: mRNA expression of PAP and NT5E in rat CP.Taste buds were isolated from digested epithelial tissues of rat CP by means of a patch pipette (A). Expression of Pap and Nt5e was examined by nested PCR (B). Ntpdase2, Plcb2, Gust, Aadc and Kcnq1 served as positive controls for taste buds including taste cells. Taste buds, CP and N.C. denotes taste buds that were isolated with a patch pipette (arrowhead, A), taste bud-containing epithelium in rat CP and negative control (H2O), respectively.

Fig 5: Immunohistochemical localization of NT5E in transverse sections of rat CP.Representative photomicrographs for double staining with NT5E (A, green; B and C, red), and the type I cell marker NTPDase2 (A, red), the type II cell marker PLC-ß2 (B, green), or the type III cell marker SNAP25 (C, green) are shown. In panel A, double staining for NT5E and NTPDase2 was performed directly using Alexa Fluor®-conjugated primary antibodies, as described under Materials and Methods. The nuclei were counterstained with Hoechst 33258 (blue). In the photomicrographs on the right, cryosections were treated with the first antibody-free solution, and immunoreactivity due to the second antibodies only was used as a negative control (N.C.), as shown with Alexa Fluor® 488 and/or 594. Scale bar, 50 µm.