Fig. 5.

Lipid accumulation and models interaction of HSP70 with oxidized LB. a–d DCs were generated from bone marrow HPC of wild-type (WT) and CD103^−/− (KO) mice using 9 day culture with GM-CSF and FLT3L followed by 48 h incubation with TES. a Phenotype of cells demonstrating lack of CD103 expression in CD11c⁺ DCs. b Lipid level in DCs after staining with BODIPY in DC treated with TES. c LB (BODIPY staining) in sorted TES-treated CD11c⁺CD172⁻ DCs analyzed by confocal microscopy. Typical example of staining (blue—DAPI, green—BODIPY) and the proportion of DCs with large LB (>0.4 µm) calculated per cell are shown. The number of cells counted is shown on the graph. Scale bar = 50 µm. d Cross-presentation of long OVA-derived peptide by DCs. *p < 0.05, ***p < 0.001 (unpaired two-tailed Student's t-test) from WT. e Volcano plot showing lipidomics analysis of oxidized TAG (singly- and doubly-oxygenated TAG, blue) and oxidatively truncated TAG (red) in TES-DC vs. Ctr-DC Horizontal dashed-line corresponds to the significance level of p = 0.05 with points above the line having p < 0.05 and points below the line having p > 0.05. f The amount of various ox-tr-TAG molecular species in control DC and DC treated with TES. g CG-MD simulations of density of POPC, non-oxidized TAG and oxidized lipids in the LB along the z-axis of the simulation box. h 3D confocal analysis of co-localization between HSP70 and LB in BM derived DC treated with TES. Green—LB, red—HSP70