Fig. 2.

Apical domain location of endogenous NHE regulatory factor (NHERF)1 and NHERF2 in Caco-2/bbe cells. z-Sections of Caco-2/bbe monolayers infected with 2.0 × 10¹⁰ particles/ml were immunostained with anti-HA monoclonal antibodies and NHERF1 or 2 polyclonal antibodies. NHERF1 and NHERF2 were localized to the apical pole, with NHERF1 demonstrating more overlap with NHE3 than NHERF2. The same settings (laser power, gain) were used for images in A–C. In D, pairs of images are shown, with images on the left representing staining using the same gain settings as A–C and images on the right shown with increased gain in order to demonstrate the different location of NHERF2 compared with NHERF1 and NHE3. Scale bars, 10 μm (A, C) and 2.5 μm (B, D). z-Series images were captured with a ×63 water immersion objective, and images were reconstructed with MetaMorph software; x–z, y–z, and x–y sections are shown in A and C and x–z sections in B and D.