FIGURE 1.

Detection of EPI64 subfamily members in mouse pancreas and isolated pancreatic acini by RT-PCR and Western blotting. A, RNA was extracted, and the mRNA levels of EPI64, 64B, and 64C were examined by RT-PCR for intact pancreas and acini. Two pairs of primers (P1 and P2) covering different regions of the mRNA were used for each gene. cDNA from mouse brain or spleen was used as positive control; β-actin serves as internal control. B, total lysates obtained from different mouse tissues and anti-TBC1D10B antibody were used for Western blotting. 293 cell lysates were used as a positive control. C, EPI64 and EPI64B immunofluorescence was localized in isolated mouse pancreatic acini. Mouse pancreatic acini were cultured with adenoviruses encoding HA-tagged EPI64 (A, C, and E) or EPI64B (B, D, and F) for 16 h after isolation. Acini were fixed and cryosectioned. Immunofluorescence was performed using anti-HA antibody.