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Figure 3. CRIM1 is required for junctional localization of E- and C-cadherin in the neural plate. (A) Immunofluorescence labeling of whole-mount Xenopus embryos after injection of translation-blocking XLCAB MOs. Embryos were co-injected with mRNA encoding GFP at the 4-cell stage and were fixed and labeled at stage 13 (early neurula) with antibodies to GFP (green), E-cadherin (A, C, D, G, H, red) or C-cadherin (B, E, F, I, J, red). Cadherin junctional complexes were visualized by combining multiple optical sections generated by confocal microscopy. In lower magnification images (C, D, E, F) it is apparent that tracer positive regions have lower levels of cadherin immunoreactivity and are irregularly shaped. In the magnified regions (G, H, I, J) indicated by white corner marks in (C, D, E, F) the loss of cadherin immunoreactivity in tracer positive cells is more obvious. The gray line between panels indicates separated color channels of the same image. (K) Graphs show the measured average E-cadherin (K) and C-cadherin (L) junctional staining intensity between two tracer-negative, one tracer negative and one tracer positive, or two tracer-positive cells (n = 20 pairs for each categories).
doi:10.1371/journal.pone.0032635.g003 |