Figure 5. Cell autonomous loss of TJ function and aPKC localization in the absence of Merlin in vitro.
(A) Confocal imaging showing the colocalization of ZO-1 and E-cadherin to PAs in both scr-shRNA and Nf2-shRNA-expressing PAM212 keratinocytes prior to calcium stimulation. After calcium stimulation ZO-1 and E-cadherin localize to distinct apical junctional rings. In contrast, ZO-1 and E-cadherin continue to colocalize to punctate structures in calcium-stimulated Nf2-shRNA-expressing cells. XZ sections are shown at the bottom of each panel. Nuclei labeled with DAPI (blue). Bars, 20 μm.
(B, C) Trans-epithelial resistance (TER) was measured across confluent control (black) or Nf2-shRNA-expressing (red) PAM212 keratinocyte (B) or Caco-2BBe intestinal epithelial cell (C) monolayers that had been stimulated with calcium for the indicated times. Values = mean +/− SD.
(D, E) Immunostaining reveals that aPKC (green) localizes primarily to cell:cell boundaries in calcium-stimulated wild-type keratinocytes but not in K14-Cre;Nf2lox/lox keratinocytes. Nuclei are stained with DAPI (blue). Bars, 15 μm.