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. Author manuscript; available in PMC: 2011 Jan 1.
Published in final edited form as: Nat Med. 2010 Jun 6;16(7):821–827. doi: 10.1038/nm.2167

Figure 4.

Figure 4

Efficient epidermal-specific lentivirus RNAi-mediated knockdown of Ctnna1 faithfully recapitulates phenotypic abnormalities shown by K14-Cre conditional and LV-Cre induced knockout counterparts. (a) TRC RNAi library shRNA constructs (arrowheads) corresponding to Ctnna1. Numbers correspond to TRC nomenclature. (b) Anti–α1-catenin and GAPDH immunoblots of protein lysates of cells FACS-sorted from embryos infected with LV-GFP harboring Ctnna1-specific shRNAs (shCtnna1) and control scrambled shRNA (shScram). (c) Quantification of α1-catenin levels from blot in b. (d–i) Comparative analyses of representative P0 skin sections of conditional Ctnna1 knockout (Ctnna1 cKO), shCtnna1-912 knockdown (Ctnna1 RNAi) and shScram control (Scram RNAi) embryos. (d) α1-Catenin immunolabeling reveals efficient knockdown in all shCtnna1-912 infected (H2B-GFP+) but not uninfected cells. (e–g) Morphological and adherens junction defects, not found in shScram RNAi–infected skin, are similar between cells infected with Ctnna1 cKO (f) and Ctnna1 RNAi (g). Boxed areas are shown in insets. Arrowheads denote hair follicles derived from infected epidermis. Note that asymmetric E-cadherin localization seen in shScram RNAi–infected (e) or uninfected areas (f) is consistently lost in Ctnna1 RNAi–infected skin (g), indicative of a planar cell polarity defect. (h,i) Suprabasal keratin 6 (K6), often reflective of enhanced basal cell proliferation, is detected in Ctnna1 cKO (h) and Ctnna1 RNAi (i) cell clones. Transduced cells are identified by their YFP or H2B-GFP expression. Nidogen (Nido) marks the basement membrane and dermal blood vessels. Epidermal adherens junctions are marked by antibody to E-cadherin (Ecad). Primary antibodies are noted on each frame, with color coding according to secondary antibodies used. Scale bars, 50 μm.