Fig 1: Increased Dvl2–EGFP localization at the plasma membrane in Cftr KO crypt-base columnar cells. (A) Representative images of WT/Dvl2-EGFP/RosamT/mG (top) and Cftr KO/Dvl2-EGFP/RosamT/mG (bottom) enteroid crypts showing Dvl2-EGFP (green), RosamT/mG (red), and merged images. Magnified merged and differential interference contrast merged with Dvl2–EGFP (DIC+Dvl2–EGFP) images of crypt base (from white boxes), were magnified (×3.6). P, Paneth cell; L, crypt lumen. White dashed lines outline apical membrane orient toward the crypt lumen. (B) Cumulative pHi data for the supranuclear region of CBCs in WT and Cftr KO enteroids. *P < .03 vs WT; n = 4–8 crypts from 4 WT and Cftr KO matched mice. (C) Crypt base model depicting the measurement by confocal microfluorimetry of Dvl2–EGFP intensity 2 pixels interior to the lateral-Lateral Membrane (lateral to central crypt axis) and medial-Lateral membrane (medial to central crypt axis) plasma membrane of CBCs. Green dashed line, central crypt axis. Brackets indicate sites of measurement on the supranuclear lateral plasma membranes. Paneth, Paneth cell. (D) Left: Cumulative data showing the average Dvl2–EGFP intensity ratio within 1.15 µm of the CBC supranuclear plasma membrane of individual mice (small circles) and overall average (large circles) for WT/Dvl2-EGFP/RosamT/mG (WT) and Cftr KO/Dvl2-EGFP/RosamT/mG (Cftr KO) sex-matched mouse pairs (n = 7 and 6 mice, respectively). Dashed line indicates the average Dvl2–EGFP pixel intensity for the entire supranuclear region of CBC, which has been set to 1.0 (average intensity = 59.5 ± 8.8 for WT and 88.5 ± 15.9 for Cftr KO; ns; n = 7–6 mice, respectively; average of 1–3 CBCs/crypt from 2 to 4 crypts of passage 1 and 2 enteroids). *P < .01 vs WT plasma membrane. Right: Cumulative data showing the average Dvl2–EGFP intensity ratio at the individual supranuclear lateral-Lateral and medial-Lateral plasma membranes of individual mice (small circles) and overall average (large circles) for WT/Dvl2-EGFP/RosamT/mG (WT) and Cftr KO/Dvl2-EGFP/RosamT/mG (Cftr KO) sex-matched mouse pairs (n = 7 and 6 mice, respectively). Dashed line indicates the average Dvl2–EGFP pixel intensity for the entire supranuclear region of CBC, which has been set to 1.0. +P < .002 vs WT lateral-Lateral cell membrane; *P < .001 vs WT medial-Lateral cell membrane. (E) Immunofluorescence image of Fz7 (cyan), plasma membrane (red), and merged (light yellow) of CBCs in WT and Cftr KO enteroid crypts. Representative of 3 WT–Cftr KO mouse pairs. L, lumen. White arrowheads denote lack of Fz7 staining at apical membrane (red).