Apoptosis Assay in Min6 Cells

A Caspase-Glo 3/7 (Caspase-Glo®, Promega) apoptosis detection kit was used to analyse Min6 cell apoptosis as described previously [7 (link)]. Groups of 2 × 10⁴ Min6 cells were maintained in culture in DMEM containing 2% FBS in the absence or presence of exogenous Wnt5a (0.05 μg/ml) for 48 h and were subsequently incubated for 20 h in the absence or presence of a cytokine cocktail (1 U/μl TNF-α, 0.05 U/μl IL-1β, and 1 U/μl IFN-γ) to induce apoptosis. Apoptosis was assessed by measuring caspase-3/7 activity as described previously [7 (link)].

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Xu W., Jones P.M., Geng H., Li R., Liu X., Li Y., Lv Q., Liu Y., Wang J., Wang X., Sun Z, & Liang J. (2020). Islet Stellate Cells Regulate Insulin Secretion via Wnt5a in Min6 Cells. International Journal of Endocrinology, 2020, 4708132.

Publication 2020

(Caspase-Glo®,

Apoptosis Caspase Caspase 3 Caspase 7 Cells Cytokine Ifn γ Il 1β Promega Tnf α Wnt5a

Corresponding Organization : Zhongda Hospital Southeast University

Other organizations : King's College London

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Variable analysis

independent variables

Presence or absence of exogenous Wnt5a (0.05 μg/ml)

dependent variables

Caspase-3/7 activity (apoptosis)

control variables

Cell type (Min6 cells)
Culture medium (DMEM containing 2% FBS)
Incubation time (48 h for Wnt5a treatment, 20 h for cytokine treatment)

controls

Negative control: Min6 cells cultured in the absence of Wnt5a and cytokines
Positive control: Min6 cells cultured in the presence of a cytokine cocktail (1 U/μl TNF-α, 0.05 U/μl IL-1β, and 1 U/μl IFN-γ) to induce apoptosis

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