Fig. 6: Adhesion-incompetent E-cadherin mutant interacts with a specific pool of actin filaments.
From: Actin-dependent α-catenin oligomerization contributes to adherens junction assembly
a Representative image of Ec/PcKO-A431 cells expressing WK-EcGFP stained for GFP (green) and for F-actin (red). Left: projections of all optical z slices show a broad localization of the mutant obscuring its plasma membrane organization. Bar, 10 µm. Right: a single optical z slice of the ventral membrane of the left image. A white dashed box of the left image is deconvolved and zoomed in the second row (z-slice of ventral membrane, deconvolved). Projection of five z slices (spanning 1 µm of the apical cell region) of a yellow box shown on upper image is zoomed on the bottom. Bars, 5 µm. Note that WK-EcGFP is enriched in the apical raffles, invadopodia (yellow arrow), and cell-cell contacting protrusions (white arrow). b The line scan performed along the dashed line shown in (a) shows the mutant in invadopodia (Inp) and in protrusion (Prtr). c WK-EcGFP cells were imaged for GFP (GFP, green) and myosin-1c (myo1c, red). Only an optical z slice with ventral membrane is shown. Bar, 10 μm. The boxed area is zoomed on the bottom. Bar, 5 μm. The optical XZ cross section along the dashed line is shown in (e). d Projections of all z slices of the EcGFP-expressing Ec/PcKO-A431 cells stained for GFP (green) and myosin 1c (red). Bar, 15 μm. The boxed area is zoomed on the bottom and presented as all z projections (All z slices) or only as a slice of ventral membrane (Z-slice of ventral membrane). Bar, 10 μm. f The line scan of GFP (green) and myosin-1c (red) fluorescence along the dashed lines shown in (c) and (d) (A.U., arbitrary units). Note that myosin-1c co-localizes with WK-EcGFP but is separated from EcGFP in AJs. g Average Pearson’s correlation coefficient between GFP and myoin-1c at cell-cell contacts in WK-EcGFP- and EcGFP-expressing cells. Correlations were calculated from single z slices of 15 areas (from 5 representative images) of the ventral membrane. The means +/- SD are indicated by bars. A two tailed Student’s t-test was used. ****P < 0.0001.
