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. Author manuscript; available in PMC: 2016 May 17.
Published in final edited form as: Immunity. 2015 Nov 10;43(5):923–932. doi: 10.1016/j.immuni.2015.10.009

Figure 4. Activation of caspase-1 leads to pannexin-1 cleavage and ATP release.

Figure 4

(A–C) BMMs were primed overnight with LPS and then transfected with ultrapure S. minnesota RE595 LPS packaged with DOTAP.

(A) Immunoblotting for pannexin-1 before and after LPS transfection in wild-type (C57BL/6), Casp11−/− and Panx1−/− BMMs.

(B) Pannexin-1 cleavage after LPS transfection in Casp11−/− BMMs reconstituted with lentiviral vectors expressing wild-type caspase-11, caspase-11 C254S and vector alone. Immunoblotting for caspase-11 and β-actin are also shown.

(C) ATP release (upper panel), cytotoxicity (middle panel) and pannexin-1 cleavage (lower panel) were measured after LPS transfection in Panx1−/− BMMs reconstituted with wild-type pannexin-1, pannexin-1 D378A lentiviruses or control vector. Immunoblotting for pannexin-1 and caspase-11 in wild-type BMMs is also shown as a control.

(A–C) Data are representative of at least 3 experiments. Graphs show mean ± SD of triplicate cultures, * p <0.05. See also Figure S6.