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. 2018 Aug 6;128(9):3840–3852. doi: 10.1172/JCI120888

Figure 5. Impaired lysosomal degradation and increased surface expression of MHC II molecules in autophagy-deficient M-MDSCs.

Figure 5

(A) Heat map of differentially expressed genes in M-MDSCs isolated from spleens of B16-F10–inoculated Atg5ΔLysM and control mice (n = 3 mice per group). (B) Heat map of differentially expressed genes related to the lysosomal function in M-MDSCs isolated from the spleens of B16-F10 inoculated Atg5ΔLysM and control mice (n = 3 mice per group). (C) MFI of lysosensor in M-MDSCs from spleen (*P = 0.0470) and tumor (**P = 0.0335) of B16-F10–inoculated Atg5ΔLysM and control mice (n = 5 mice per group). (D) Percentage of protein degradation, using [3H] leucine, in M-MDSCs isolated from the spleens of B16-F10 inoculated Atg5ΔLysM and control mice treated with lysosomal inhibitors (NH4Cl and leupeptin or bafilomycin) or left untreated (n = 3 mice per group). *P = 0.0134, **P = 0.0084, ***P = 0.0195, ****P = 0.0128, #P = 0.0179, ##P = 0.0088, ###P = 0.0264. (E) Representative histograms for the expression of IAb by M-MDSCs of spleen or tumor of Atg5ΔLysM and control mice, n = 5 mice per group. (F) Representative confocal microscopy images for LAMP-1 (red), IAb(green), DAPI (blue), and Pearson’s correlation of IAb versus LAMP-1 (***P < 0.0001) in sorted M-MDSCs from splenocytes of B16-F10–inoculated Atg5ΔLysM and control mice (n = 4 mice/group). Scale bar: 10 μm. (G) Representative histograms for the expression of IAb by M-MDSCs isolated from spleens of B16-F10–inoculated Atg5fl/fl mice after in vitro stimulation with TES in the presence of NH4Cl or chloroquine. Geometric mean of IAb (***P < 0.0001, *P = 0.048) and relative expression of Ciita and IAb (***P < 0.0001) are shown, n = 8 mice per group. Results are mean ± SEM. Statistical significance was obtained by unpaired Student’s t test, or 2-way ANOVA (D and G). Representative results from 3 independent experiments are shown.