IBEX immunofluorescence images of skin from CD11c-DOG mice treated or untreated with diphtheria toxin

Iterative Bleaching Extends Multiplexity (IBEX) immunofluorescence was performed on skin samples collected from CD11c-DOG mice either treated with diphtheria toxin (Skin_CD11cDOG_with_Diphtheria_Toxin) or phosphate-buffered saline (PBS) treated controls (Skin_CD11cDOG_no_Diphtheria_Toxin). For dendritic cell depletion, CD11c-DOG mice received three systemic intravenous injections of diphtheria toxin (16 ng/g body weight) on days −3, −2, and −1. Control mice PBS following the same schedule. In addition, a local subcutaneous injection of diphtheria toxin (16 ng) was administered on day −1 at the site designated for tissue collection. Mice were euthanized on day 0 in accordance with national and international regulations for animal care and use. A square section of flank skin was excised from each mouse at the site of the last intradermal injection. Subcutaneous adipose tissue was mechanically removed using forceps. Skin samples were subsequently processed according to the 2D Manual IBEX protocol for multiplex immunofluorescence imaging.