Figure 3: ChA activates effector caspases but induces caspase-independent cell death of M2.
From: Modulation of actin dynamics as potential macrophage subtype-targeting anti-tumour strategy
(A) PARP cleavage and (B) caspase-3/7 activation in macrophages after treatment with vehicle (0.1% DMSO) or ChA for (A) the indicated times or (B) 24 h. ChA or staurosporine (stsp) were added 30 min prior to polarization. Pictures shown in (A) are representative Western blots for cleaved PARP and GAPDH (for normalization) of three independent experiments. In (B), caspase-3/7 activation was determined by using Apo-ONE Homogenous Caspase-3/7 assay; data are means + SEM; n = 3. (C) PARP cleavage, (D) LDH and (E) MTT assays of M2 pre-treated (30 min, 37 °C) with vehicle (0.1% DMSO), the pan-caspase inhibitor QVD, or the caspase-1 inhibitor YVAD (10 μM, each), and incubated with vehicle (0.1% DMSO) or ChA for (C,D) 24 h or (E) 48 h. ChA was added 30 min prior to polarization or corresponding incubations without stimuli (for M0). Western blots for cleaved PARP or GAPDH shown in (C) are representative of three independent experiments. In (D,E) data are expressed as percentage of (D) Triton X-100 (1%) or (E) vehicle controls, means + SEM; n = 3; *p < 0.05, **p < 0.01, ***p < 0.001 vs vehicle control.
