Figure 3. Results of the immunocytostaining analysis using anti-V5 antibody for the HCE-T cells transfected with expression vector harboring the wild-type or mutated TACSTD2 gene tagged with V5-epitope. Immunolocalization at the plasma membrane is apparent in the HCE-T cells transfected with the wild-type (A) TACSTD2. In HCE-T cells transfected with the mutated (B: p.Ile281SerfsX23, C: p.Tyr225X, D: p.Gln118X) TACSTD2, immunoreactivity was observed not at the plasma membrane but in the cytoplasm with slightly intensified signal around their nucleus. In the HCE-T cells transfected with wild-type TACSTD2 (E and F), no apparent change was observed by the detergent treatment (0.1% Tween-20 for 30 min; F). However, in the HCE-T cells transfected with mutated TACSTD2 (G and H: p.Ile281SerfsX23, I and J: p.Tyr225X), detergent treatment (H, J) significantly increased the number of the immunopositive cells as compared to those with no detergent treatment (G and I).