Fig. 4

Interaction between LB and HSP70 in DC. a Confocal microscopy analysis of LB in DCs treated with TES. Typical example is shown. Scale bar = 10 µm. Proportion of DCs containing more than three large LB (minimal size 0.4 µm) was calculated. For each sample at least 100 cells were counted. The results from three samples are shown. **p < 0.01. b, c Confocal microscopy of LB and HSP70 in DC treated with TES (b) or in human monocyte derived DC treated with TCM (c). Scale bar = 10 µm. Four different experiments were performed and typical example is shown. The proportion of DCs with co-localization of LB and HSP70 b, c was calculated from the total cells containing LBs. At least 50 cells in each sample were counted. Four samples were evaluated d. DCs were generated from HPC using GM-CSF and FLT3L and then exposed to TES. d Confocal microscopy of sorted CD103⁺ DCs loaded with long peptide. Scale bars = 50 µm. e CD103⁺ and CD103⁻ DCs were sorted from dLNs of TB mice and evaluated. On the left—typical example of staining. On the right—proportion of co-localization of lipid bodies with HSP70. Scale bar = 10 µm. Four independent experiments were performed. ***p < 0.001 in unpaired two-tailed Student's t test. f Co-localization of HSP70 with LB in DC sorted from draining LN of LLC bearing mice or LN of naïve mice. Left—typical examples of staining. Scale bar = 1 µm. Right—number of cells with co-localization LB and HSP70 out of 100 cells counted. Cumulative results obtained from three mice. p value in Fisher’s rank test. g Co-localization of HSP70 and CD11c in DC treated with TES. Left—typical example of staining. Scale bar = 10 µm. Right—% of co-localization for HSP70 and CD11c in gated part of the cell calculated using Leica Software. At least 40 cells were counted in each sample, total four samples