ATG5-deficient Langerhans cells present endoplasmic reticulum swelling but no unfolded protein response. (A) MFI for ER-tracker on epidermal LCs treated or not with Wortmannin. Data are pooled from at least three independent experiments, with each point representing one individual 3-wk-old mouse. Statistical analysis: Mann–Whitney U test (*, P < 0.05). (B) Representative half-set overlay (left panel) and MFI (right panel) of ER-tracker in epidermal LCs of control (Atg5^WT and Atg5^WT/Δ) and Atg5^ΔCd207 mice. Data are pooled from at least three independent experiments, with each point representing one individual 3-wk-old mouse. Statistical analysis: Kruskal–Wallis one-way ANOVA followed by Dunn’s multiple comparison test (**, P < 0.01). (C) Representative immunofluorescent staining of the endoplasmic reticulum using the ER-tracker dye on epidermal LCs of 3-wk-old Atg5^WT (Video 3) and Atg5^ΔCd207 (Video 4) mice. CD207: green; ER-tracker: red; DAPI: blue. Scale bar: 10 µm. (D) Expression of Ern1, total and spliced Xbp1, and Ddit3 mRNAs, in epidermal LCs of 3-wk-old control (Atg5^WT and Atg5^WT/Δ) and Atg5^ΔCd207 mice. Fold changes were calculated relative to mRNA expression in cells of Atg5^WT control mice. Data are pooled from at least three independent experiments, with each point representing one individual mouse. Statistical analysis: Kruskal–Wallis one-way ANOVA followed by Dunn’s multiple comparison test (ns, P > 0.05).