Fig. 2: Direct contact of live Eh via Gal-lectin and EhCP-A1/EhCP-A4 mediates ATG16L1 protein degradation. | Mucosal Immunology

Fig. 2: Direct contact of live Eh via Gal-lectin and EhCP-A1/EhCP-A4 mediates ATG16L1 protein degradation.

From: Entamoeba histolytica exploits the autophagy pathway in macrophages to trigger inflammation in disease pathogenesis

Fig. 2

a Eh were pre-treated with 55 mM galactose for 5 min before incubation with THP-1 macrophages for 10 min at 1:20 ratio. b THP-1 macrophages were stimulated with live Eh (1:20 ratio), Eh secreted proteins (SP, 50 μg), Eh whole lysate (WL), Eh cytoplasmic component (CM), and Eh membrane component prepared from equivalent amount of Eh for 10 min. c THP-1 macrophages were incubated with native Gal-lectin (500 ng/ml) for increasing times. d Eh were pre-treated overnight with E-64 (100 μM) and incubated with macrophages for different time points along with non-treated Eh. e THP-1 macrophages were stimulated with wild type Eh, EhCP-A5 deficient Eh, and E-64 treated Eh for 10 min and 20 min. Eh were also pre-treated with specific inhibitor for EhCP-A1 (WRR483, 20 μM), EhCP-A4 (WRR605, 20 μM) individually or both together and incubated with (f) THP-1 and (g) bone marrow derived macrophages (BMDMs) for 10 mins. Equal amount of lysed cell lysates was loaded onto SDS-PAGE gel and immunoblotted with indicated antibodies. Cells without treatment is symbolized as (−ve). Eh-macrophage 1:20 ratio used. Results are representative of three independent experiments (n = 3).

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