Figure 5.

Atg16l1 deficiency in CD8α+ dentritic cells (DCs) does not induce immune tolerance and does not protect against atherosclerosis in Ldlr⁻/⁻ (low-density lipoprotein receptor–deficient) mice. A, Strategy for conventional DC (cDC) purification from the spleen of CD11c^Cre+ Atg16^flox/flox, Clec9a^Cre+ Atg16l1^flox/flox and control mice (CD11c^Cre− Atg16^flox/flox and Clec9a^Cre− Atg16^flox/flox) and analysis of Atg16l1 expression by real-time quantitative polymerase chain reaction (RT-QPCR; normalized on 36B4). n=5-6 mice/group; P<0.0001 for comparisons between all groups (Kruska-Wallis); post hoc uncorrected Dunn’s test, ***P=0.0003 DC CD8α⁺ CD11c^Cre+ Atg16^flox/flox; ***P=0.0008 DC CD8α⁺ Clec9a^Cre+ Atg16^flox/flox vs DC CD8α⁺ control; *P=0.056 DC CD11b⁺ CD11c^Cre+ Atg16^flox/flox vs DC CD11b⁺ control. B, Representative flow chart showing the distribution of CD8α⁺ and CD11b⁺ DC subsets and the quantification of the proportion of cDCs in the spleens of Clec9a^Cre+ Atg16^flox/flox and control mice after bone marrow transplantation (BMT) in Ldlr^−/− mice and 8 wk of high-fat diet (HFD). C, Representative flow chart showing the distribution of CD8⁺ and CD4⁺ T cells and the quantification of the proportion of each subset in the spleens of Clec9a^Cre+ Atg16^flox/flox and control mice after BMT in Ldlr^−/− mice and 8 wk of HFD. D, Representative flow chart showing the proportion of IFNγ (interferon γ)-producing CD4⁺ T cells, and the quantification, in the spleens of Clec9a^Cre+ Atg16^flox/flox and control mice after BMT in Ldlr^−/− mice and 8 wk of HFD. E, Representative flow chart showing the proportion of CD4⁺ regulatory T cells (CD3⁺CD4⁺Foxp3 [forkhead box P]⁺CD25^high), and the quantification in the spleens of Clec9a^Cre+ Atg16^flox/flox and control mice after BMT in Ldlr^−/− mice and 8 wk of HFD. F and G, Quantification and representative pictures of Oil red O staining for atherosclerotic lesions analysis on en face thoracic aorta (F) and in the aortic sinus (G) of Clec9a^Cre+ Atg16^flox/flox and control mice after BMT in Ldlr^−/− mice and 8 wk of HFD. B–E, G, Seven to 8 mice per group. F, Six mice per group. Data are representative of 2 independent experiments. FACS indicates fluorescence-activated cell sorting; and MHC, major histocompatibility complex.