FIGURE 2:

PLEKHA5 and PLEKHA6 are localized along the basal, apical, and lateral plasma membranes in cultured cells and recruit PDZD11. (A, F–L) Immunofluorescence (IF) microscopy of endogenous PLEKHA5, PLEKHA6, and PLEKHA7 in cultured cells, using either zonular markers (ZO-1, CGN) or the AJ/lateral marker E-cadherin, or the apical marker GP135, or the microtubule protein α-tubulin as references. Different cell types and supports were used, as indicated: (A) mCCD (Transwells); (F, G) MDCK (Transwells); (H–K) MDCK (cysts in Matrigel); (L) Hap1 (coverslips). (B–E) IF microscopy of exogenous PDZD11-HA and GFP-tagged PLEKHA6 (B), PLEKHA7 (C), PLEKHA5 (D), and GFP alone (E, negative control) in mCCD cells (Transwells). (M–P) IF microscopy of endogenous PLEKHA5 (M, N) or exogenous GFP-tagged PDZD11 (O, P) in MDCK or Hap1 cells grown on coverslips, treated with either DMSO (M, O) or nocodazole (N, P), using antibodies against α-tubulin to label microtubules. For cells on Transwells (A–G), Z sections taken at the horizontal middle positions are shown below XY images. Arrows indicate labeling, arrowheads indicate low/undetectable labeling. Junctional, basal, apical plasma membrane (PM), cytoplasmic (cyt/cytopl.), subapical cytoplasmic (cyt.), and lateral (lat.) localizations/pools are indicated by arrows. Bars = 20 µm.