Figure 4.

Correct deposition of human C7 at the DEJ in immunodeficient mice after intradermal injection of ABE- or PE-treated primary RDEB fibroblasts

(A) Scheme of the experiment in which ABE-/PE-treated patient-derived fibroblasts were injected into the mouse model. (B) Immunofluorescence staining to visualize the C7 protein (row 1). NHDFs, non-edited fibroblasts from Pat1, ABE-treated RDEB fibroblasts, PE-treated RDEB fibroblasts (5 × 10⁶ cells/150 μL of phosphate-buffered saline), or phosphate-buffered saline were intradermally injected into the back skin of immunodeficient mice. Two weeks after the injections, immunofluorescence analysis of C7 (green) was performed using a rabbit polyclonal antibody that recognizes human C7. Immunofluorescence staining using an antibody specific for human vimentin (red) revealed the presence of human fibroblasts injected into the mouse skin (row 2). Phosphate-buffered saline-injected mouse skin was used as a negative control. The mouse experiments were repeated two times, and at least three mice were included in each cell group each time. Images of a representative mouse experiment are displayed (n = 3 repeats of each cell group in two independent experiments). The white dotted line indicates the DEJ. White arrows indicate human C7 deposited at the DEJ. Scale bars, 50 μm. Epi, epidermis; Der, dermis; Fb, fibroblasts; NHDFs, normal human dermal fibroblasts.