Figure 5.

CLL cells are unable to form the ripoptosome in response to Compound A. (a–c) IP of caspase-8 in Kym-1 and CLL cells co-cultured with 3T40 cells (a and b) or not (c) (numbers refer to CLL cases in Table 1), untreated or treated with Compound A, followed by detection of ripoptosome components caspase-8, RIP-1, FADD and cFLIP_L. CLL cells were co-cultured on 3T40 cells for 96 h, collected and treated with 100 nM Compound A for 4 h (a) or 24 h (b) or directly treated with 500 nM Compound A for 4 h (c). Kym-1 cells were exposed to 20 nM Compound A for 4 h. Caspase-8 IP was performed from total lysates. (a) Pre-IP, post-IP and IP samples were prepared, and western blotting was performed for caspase-8 and RIPK1. Actin serves as a loading control. (b) Pre-IP and IP samples were prepared, and western blotting was performed for caspase-8, RIPK1, FADD and cFLIP_L. Actin serves as a loading control. *marks an aspecific band. (c) Pre-IP and IP samples were prepared, and western blotting was performed for caspase-8 and RIPK1. Actin serves as a loading control