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. Author manuscript; available in PMC: 2019 May 15.
Published in final edited form as: J Immunol. 2018 Apr 2;200(10):3556–3567. doi: 10.4049/jimmunol.1701504

Figure 1. Phagocytosis is required for CS-induced LTB4 and IL-1β production.

Figure 1

BMDMs from WT mice (3 × 106 cells) were cultured O/N in 12 well dishes, LPS-primed and stimulated with 100 μg/cm2 CS for 6 h in the presence or absence of Cytocholasin D (CytD, 10 μM) as described in methods. ELISA was performed to assess levels of (a) LTB4 and (b) IL-1β in the culture supernatants. (c) Mouse bone marrow-derived mast cells, mouse bone marrow-derived neutrophils and human neutrophils were stimulated with 100 μg/cm2 CS in the presence or absence of CytD as described in methods. Levels of LTB4 production was measured using ELISA. Data is representative of one of the three experiments. Data are expressed as mean ± SEM. *p< 0.05, **p< 0.01, ***p< 0.001 non-parametric t-test. (d) CS uptake in the presence and absence of CytD was analyzed 2h post stimulation with 100 μg/cm2 CS in LPS-primed BMDM. Cell membrane was stained with Alexa fluor 594 Cholera toxin Subunit B (red), nucleus with DAPI (blue) and CS particles were observed using reflective microscopy. Representative images shown are from one of the three independent experiments.