Fig. 7: Glucose uptake activates mTOR and pro-inflammatory responses in EV-recipient cells.
From: Tissue-infiltrating macrophages mediate an exosome-based metabolic reprogramming upon DNA damage
a Immunofluorescence detection of iNOS accumulation (indicated by the arrowhead) in PPCs exposed to culture media (CM) and the EVs derived from MMC-treated and untreated control macrophages (n = 3, >600 cells per treatment) or b Er1F/− and Er1F/+ macrophages (n = 3, > 1200 cells/treatment) (see also Supplementary Fig. 8D). c Immunofluorescence detection of NF-kβ nuclear translocation (indicated by the arrowhead) in PPCs exposed to CM and the EVs derived from MMC-treated and untreated control macrophages or (n = 3, >700 cells per treatment). d Er1F/− and Er1F/+ macrophages (see also Supplementary Fig. 8E). (n = 3, >500 cells per treatment). e Immunofluorescence detection of NF-kβ (shown by the arrowhead) in PPCs exposed to CM supplemented with EVs from MMC-treated and control macrophages upon low (5 mmol) or high (15 mmol) glucose concentration (n = 4, >750 cells per treatment; see also Supplementary Fig. 9A–C). f Western blotting of phosphorylated pS6K, phosphorylated p4EBP1, phosphorylated pAKT1, phosphorylated pAKT2 and phosphorylated pAKT protein levels in Er1F/− and Er1F/+ pancreata (n = 4). The graph represents the fold change (F.C) of indicated protein levels in Er1F/− pancreata to wt. controls. g Western blotting of pS6K, p4EBP1 and iNOS protein levels in MMC-treated macrophages exposed to rapamycin (n = 3, Rap/cin; as indicated). The graph represents the fold change of protein levels in MMC-treated macrophages exposed to rapamycin compared to MMC-treated macrophage control (ctrl.) cells. h Immunofluorescence detection of NF-kβ and 2-NBDG in PPCs (indicated by the arrowheads) exposed to CM supplemented with EVs derived from Er1F/− and Er1F/+ BMDMs in the presence or absence of rapamycin (Rap/cin). (n = 6, > 1000 cells counted per treatment). The graph shows the percenatge of positively stained cells. Error bars indicate S.E.M. among replicates (n ≥ 3). Asterisk indicates the significance set at p-value: *≤0.05, **≤0.01 (two-tailed Student’s t-test). Gray line is set at 5 μm scale.
