Fig. 4: CBRs-induced autophagy drives metabolic reprogramming and human tolerogenic DCs generation.

a Gene expression levels of hmoDCs stimulated with LPS (0.1 µg/mL) plus WIN55212-2 (WIN, 10 µM) or in the presence of the autophagy inhibitor 3-methyladenine (3-MA, 25 µM) relative to LPS-stimulated condition (n = 5). b Cytokines produced by naïve CD4⁺ T cells primed by hmoDCs stimulated in the indicated conditions after 5 days (n = 6). c Increment in FOXP3⁺ Tregs generation relative to LPS-stimulation condition. d Flow cytometry representative dot plots. e Quantification of the induced Warburg effect and lactate content in cell-free supernatants relative to LPS stimulation (n = 6). f Glucose consumption determined as metabolic rate of the indicated conditions (n = 6). g Fluorescence intensity of hmoDCs stained with Mito Tracker Red relative to LPS-stimulated condition (n = 6). h Left, western blot of hmoDCs stimulated with LPS, LPS/WIN55212-2 or LPS/WIN55212-2 in the presence of CB1 (Rimonabant, (RIM), 10 µM) and PPARα (GW6471, 25 µM) antagonists for 18 h. Right, quantification of the reactive bands by scanning densitometry. i Quantification of the induced Warburg effect, metabolic rate and fluorescence intensity of Mito Tracker Red stained cells in hmoDCs relative to LPS stimulation (n = 6). Values are mean ± SEM. Statistical significance was determined using One-way Anova. *P < 0.05, **P < 0.01.