Fig. 1

Anln depletion in the developing epidermis disrupts tissue integrity. A Wild-type keratinocytes were cultured in media containing 50μM or 1.5mM calcium, fixed, and labeled for ANLN and E-cadherin. Scale bar = 20 μm. B Whole-mount immunofluorescence images of Anln-GFP-transduced E16.5 embryos immunolabeled for E-cadherin and imaged at the middle of the basal layer. Arrows denote nuclear localization, arrow heads denote cytoplasmatic, and asterisk denotes cortical localization. Scale bar = 20 μm. C Quantitative PCR analysis of Anln mRNA in primary mouse keratinocytes transduced with scrambled shRNA (Ctrl) or one of two Anln-specific shRNAs (926 and 2981). Data are the mean ± SD of four preparations. *, P < 0.0001 (Scr vs. Anln-926); *, P < 0.0001 (Scr vs. Anln-2981) by unpaired t-test. D Western blot analyses of primary mouse keratinocytes transduced with Scr, Anln-926, or Anln-2981 shRNAs. Blots were probed with antibodies to ANLN and GAPDH (loading control). E Stereomicroscopic images of E18.5 embryos infected on E9 with shScr;H2B-GFP (Ctrl) or shAnln-926;H2B-GFP lentiviruses. Arrows denote defects in the head, tail, and limbs. Scale bar = 2 mm