Fig. 2. Tamoxifen injection into CD98hc^ΔCX3CR1 animals leads to the excision of CD98hc in monocytes and macrophages.

Following intraperitoneal tamoxifen injection, monocytes, and macrophages were isolated from the colonic lamina propria (cLP) of CD98hc^ΔCX3CR1 animals at indicated time points and analyzed for CD98hc expression by flow cytometry. a Percentage of CD98hc⁺ monocytes and macrophages (n = 3). b Histogram plots showing CD98hc staining intensity in cLP monocytes and macrophages. c Mean (±SD) percentage of CD98hc⁺ CD11c^neg, and CD11c^pos liver myeloid cells and (d) Langerhans cells (n = 3). e Histogram plots of CD11c^neg and CD11c^pos liver myeloid cells and Langerhans cells isolated from liver and epidermis, respectively. Red histograms display FMO controls, blue histograms CD98hc⁺ cells; Numbers in histograms show the percentage of CD98hc⁺ cells (b, e). The data are shown as the mean (± SD), and the results were analyzed by two-way ANOVA followed by Sidak’s correction; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 (a–d). f After gating on CX3CR1/YFP⁺ or CX3CR1/YFP⁻ CD11c⁺ or CD11c⁻ liver myeloid cells, Zeb2 expression by CD11c⁺ or CD11c⁻ liver myeloid cells were analyzed. Numbers in dot plots indicate the percentage of positive cells. g Percentage of CX3CR1/YFP⁺ or CX3CR1/YFP⁻, and percentage of Zeb2⁺ CD11c⁺ or CD11c⁻ liver myeloid cells. Each dot represents one animal; the mean is indicated. The data were analyzed by Mann–Whitney U test; *p < 0.05. Experiments were performed once with three biological replicates for CD98hc silencing kinetics, and once with four biological replicates for Zeb2 expression.