Figure 5.
TNFR2 promotes recruitment of RIP1 to TNFR1. (A) Endogenous TNFR1 immunoprecipitation (IP) in WT and TNFR2ko R-03 (responder) PDX cells treated with birinapant, as indicated. (B) Lysates and TNFR1 immunoprecipitation of sensitive (R-03, IC50 < 100 nM) and resistant (VHR-01, IC50 > 1000 nM) ALL. (C) WB in nonreducing nondenaturing conditions for TNFR1 monomers (mono), dimers, and trimers (Tri) in responder and nonresponder PDX (left panel), and WT and TNFR2ko R-03 PDX cells (right panel) treated as indicated. (D) Analysis of RIP1 phosphorylation at serine-166 (pRIP1) in WT and TNFR1ko ALL samples. (E) Lysates and caspase-8 (Casp8) IP (ripoptosome) for WT and TNFR2ko (R2ko) R-03 and VHR-10 PDX cells treated with birinapant, as indicated. Data are representative of 3 independent experiments. Bir/bir, birinapant; 10′, 10 minutes; 30′, 30 minutes.

TNFR2 promotes recruitment of RIP1 to TNFR1. (A) Endogenous TNFR1 immunoprecipitation (IP) in WT and TNFR2ko R-03 (responder) PDX cells treated with birinapant, as indicated. (B) Lysates and TNFR1 immunoprecipitation of sensitive (R-03, IC50 < 100 nM) and resistant (VHR-01, IC50 > 1000 nM) ALL. (C) WB in nonreducing nondenaturing conditions for TNFR1 monomers (mono), dimers, and trimers (Tri) in responder and nonresponder PDX (left panel), and WT and TNFR2ko R-03 PDX cells (right panel) treated as indicated. (D) Analysis of RIP1 phosphorylation at serine-166 (pRIP1) in WT and TNFR1ko ALL samples. (E) Lysates and caspase-8 (Casp8) IP (ripoptosome) for WT and TNFR2ko (R2ko) R-03 and VHR-10 PDX cells treated with birinapant, as indicated. Data are representative of 3 independent experiments. Bir/bir, birinapant; 10′, 10 minutes; 30′, 30 minutes.

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