Fig. 4

ANLN loss-of-function enhances F-actin levels in the developing epidermis. A, C, E Sagittal views of 10-μm sections of dorsal skin from shScr (ctrl) and shAnln-926 transduced E14.5 (A), E15.5 (C), and E16.5 (E) embryos. Sections were stained for F-actin. B Quantification of F-actin staining intensity from the data shown in A. Horizontal bars represent the mean F-actin intensity, and circles represent individual microscopy fields. n = 4 embryos per condition. NS denotes not significant. D Quantification of F-actin staining intensity from the data shown in C. Horizontal bars represent the mean F-actin intensity, and circles represent individual microscopy fields. n = 3 embryos per condition. P = 0.0136 for Ctrl versus Anln-926 basal layer; P = 0.0254 for Ctrl versus Anln-926 suprabasal layers by unpaired two-tailed t-test. F Quantification of F-actin staining intensity from the data shown in E. Horizontal bars represent the mean F-actin intensity, and circles represent individual microscopy fields. n = 3 embryos per condition. P = 0.0123 for Ctrl versus Anln-926 basal layer; P = 0.0392 for Ctrl versus Anln-926 suprabasal layer by unpaired two-tailed t-test. G Line scan analyses from data shown in E. Asterisks denote analyzed cells. H Sagittal views of 10-μm sections of dorsal skin from wild-type E15.5 embryos treated with DMSO (Ctrl) or jasplakinolide, fixed, and immunolabeled for E-cadherin, Par3, and nidogen. Dotted lines in A, C, E, and H indicate the dermal–epidermal border. Insets in A, C, and E show the transduced cells (H2B−GFP+). Yellow insets in A, C, E, and H show magnification of boxed areas. Red lines in C and E indicate the boundary between basal and suprabasal layers. Nuclei were stained with DAPI (blue). Scale bars = 20 μm