Caspase 3/7 Activity in C. concisus Zot-Induced Apoptosis

Caspase 3/7 activity was measured in order to examine whether C. concisus Zot caused apoptosis in Caco-2 cells. Caco-2 cells were cultured on black-walled 96-well plates with transparent bottom for 2 days and then incubated with EP and EP-ZotP14UCO-S1 proteins at 50 μg/100 μl for 2 h. Caco-2 cells incubated in DPBS were used as the negative control and DMSO (5 %) treated Caco-2 cells were used as the positive control [29 (link)]. Caspase 3/7 activity was then measured using fluorescent CellEvent Caspase-3/7 Green ReadyProbes Reagent (Thermo Fisher Scientific) as per the manufacturer’s instruction [28 (link)]. The level of caspase 3/7 activity in Caco-2 cells treated with EP and EP-ZotP14UCO-S1 were expressed as folds relative to that in the negative control Caco-2 cells.

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Mahendran V., Liu F., Riordan S.M., Grimm M.C., Tanaka M.M, & Zhang L. (2016). Examination of the effects of Campylobacter concisus zonula occludens toxin on intestinal epithelial cells and macrophages. Gut Pathogens, 8, 18.

Publication 2016

CellEvent Caspase-3/7 Green ReadyProbes Reagent

Apoptosis Caco 2 cells Caspase 3 Caspase 7 Dmso Ep 100 Proteins

Corresponding Organization :

Other organizations : UNSW Sydney, Prince of Wales Hospital, Sutherland Hospital

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Variable analysis

independent variables

EP protein
EP-ZotP14UCO-S1 protein

dependent variables

Caspase 3/7 activity

control variables

Caco-2 cells cultured on black-walled 96-well plates with transparent bottom for 2 days

positive control

DMSO (5%) treated Caco-2 cells

negative control

Caco-2 cells incubated in DPBS

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