Canonical signals are non-essential for basolateral sorting of CALHM1/CALHM3 channel. (A–B) Fluorescence double-labeling of polarized MDCKII cells expressing GFP-fused CALHM1 MLYI/4A and untagged CALHM3 LMYL/4A with GFP (green) and Na+/K+ ATPase (A) or apical membrane-bound biotin (B) (red). (C–D) MDCKII epithelia expressing GFP-fused CALHM1 MLYILL/6A and untagged CALHM3 LMYLLL/6A were double-stained with GFP (green) and Na+/K+ ATPase (C) or apical membrane-bound biotin (D) (red). Upper panel, XY image. Lower panel, XZ image reconstructed at the blue line in the upper image. Api, apical membrane; Baso, basolateral membrane; Scale bars, 10 µm. Data are representative of three experiments.