Fig. 4.
Immunofluorescent validation of selected novel brush border components. CACO-2BBE cells were stained with Alexa 568-phalloidin (red) and antibodies directed against mucin-like protocadherin (MLPCDH; A and D), protocadherin-24 (PCDH24; B and E), or harmonin (C and F), which were detected with Alexa 488-conjugated secondary antibodies (green). A–C: en face views taken near the apical surface of the cell monolayer showed that all three proteins demonstrated a punctate staining pattern that was highly mosaic. All three proteins localized strongly to the apical surface of the cells, at the top of the monolayer, as viewed in the vertical sections (D–F). Scale bars = 10 μm in A–C and 5 μm in D–F. Insets show magnified views of individual cells. Box width is 20 µm in all cases.