(A and B) Immunostaining of Aqp2(green) and Rab11a (red) (A), and Aqp2(green) and GM130 (red) (B), of si-Cont and si-Pdcd10–treated mCCD_Cl1 cells transduced with lentivirus expressing Aqp2. The square panels represent xy-sections, and the smaller horizontal strips at the bottom of each square panel represents z-sections of reconstructed 3-D confocal stacks at the at the positions indicated by the yellow dashed lines in the xy planes. (C) Representative images showing the internalization of dextran (red) in si-Cont and si-Pdcd10–transfected mCCD_Cl1 cells, and si-Pdcd10–transfected mCCD_Cl1 cells treated with Erlotinib or methyl-β-cyclodextrin (MβCD). The middle square panels represent higher magnification of the selected areas. The z-sections of the cells are indicated by the dashed line at the bottom of each square panel. (D) The quantifications of the mean cellular dextran fluorescence intensity; each dot represents fluorescence intensity normalized to si-Cont (si-Cont and si-Pdcd10, n = 4; si-Pdcd10 + Erlo and si-Pdcd10 + MβCD, n = 3). (E) Immunostaining of Ezrin (green) and p-ERM (red) in the si-Cont and si-Pdcd10–treated mCCD_Cl1 cells. The middle square panels represent higher magnification of the selected areas. The square panels represent xy-sections, and the horizontal strips at the bottom of each square panel represent z-sections of reconstructed 3-D confocal stacks. (F) The quantification of mean cellular Ezrin and p-ERM fluorescence intensity; each dot represents fluorescence intensity normalized to si-Cont (n = 6 for each group of Ezrin; si-Cont of p-ERM, n = 5; si-Pdcd10 of p-ERM, n = 4). The n value represents the field of visions in different experiments. (G) The quantification of mean cellular p-ERM fluorescence intensity after normalization with total ezrin abundance. Data are presented as mean ± SEM using unpaired t test (F and G) or 1-way ANOVA (D). ***P < 0.001; **P < 0.01. Original magnification, ×630 (A and C); ×400 (B and E).