Autophagy deficiency affects neuronal interaction genes in Langerhans cells. (A) Differentially expressed genes in Atg5^WT versus Atg5^ΔCd207 LCs of 3-wk old mice: transcripts related to neuronal interactions (GO:0030031, GO:0071526, GO:0050808, GO:0045664). (B) Representative immunofluorescence microscopy image of epidermal sheets obtained from ears of 6–12-mo-old Atg5^WT and Atg5^ΔCd207 mice and stained with antibodies against β3-tubulin (neurons) and CD207 (LCs). Scale bar: 50 µm. Quantification in Atg5^WT, Atg5^WT/−, and Atg5^ΔCd207 mice: (C) Number of CD207⁺ LCs per mm². (D) Relative area of β3-tubulin staining. Data are pooled from three independent experiments, with each point corresponding to one field of view (n = 3 mice per condition, at least 10 fields per mouse were scanned). Statistical analysis: Kruskal–Wallis one-way ANOVA followed by Dunn’s multiple comparison test (**, P < 0.01; ****, P < 0.0001; ns, P > 0.05). (E) Correlation of epidermal nerve and Langerhans cell densities in 6–12-mo-old Atg5^WT, Atg5^WT/− and Atg5^ΔCd207 mice. Based on epidermal sheet staining (C and D), the number of CD207⁺ LCs per mm² (X axis) was plotted against the relative areas for β3-tubulin⁺ nerves (Y axis). Statistical analysis: Pearson correlation test. R² correlation coefficients and P values are indicated.