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. 2017 Apr 27;67(2):333–347. doi: 10.1136/gutjnl-2016-313615

Figure 7.

Figure 7

Secretory leucocyte protease inhibitor (SLPI) enhances the monocyte clearance of apoptotic neutrophils. (A–D) CD14-isolated monocytes were cultured with medium or (rh)-SLPI (0.5 μg/mL) or dexamethasone (100 nM) for 48 hours and then coincubated (4 hours) with apoptotic neutrophils (n=3 independent experiments). (A) Representative confocal microscopy images of CMTPX-labelled monocyte engulfment of apoptotic CMFDA-labelled neutrophils (original magnification ×63); merge/z-stack images: arrows showing colocalised/engulfed cells. (B) Data show Mer tyrosine kinase (MerTK) expression in monocyte subsets after culture (48 hours) with different treatments. (C and D) Representative flow cytometry plots and percentage of monocytes that phagocytosed CMFDA-labelled neutrophils. (E and F) CD14-isolated monocytes were cultured with medium or (rh)-SLPI (0.5 μg/mL) for 48 hours and then coincubated (4 hours) with apoptotic Huh-7 hepatoma cells (n=3 independent experiments). (E) (Upper) Representative confocal microscopy images of CMFDA-labelled Huh-7 cells and (lower) representative Annexin-V/7-AAD staining of Huh-7 cells treated with/without 20 μM STS (50 μm, scale bars). (F) Representative flow cytometry analysis and percentage of monocytes that phagocytosed CMFDA-labelled (STS-treated) apoptotic Huh-7 cells. Non-parametric (Mann-Whitney) statistical analysis was used. Data are expressed as median values with IQR. *p<0.05, **p<0.01, ****p<0.0001. ns, non-significant.