Birinapant, GDC-0152, embelin, and vorinostat (suberoylanilide
hydroxamic acid) were purchased from Selleck Chemicals. Concentrations of
drugs used were based upon previously published whole blood
Cmax and
Cmin data for each drug (Amaravadi et al., 2015 (link); Flygare et al., 2012 (link); Zutshi et al., 1990 (link)) and were prepared in
dimethyl sulfoxide. Corresponding volumes of DMSO
(5μL DMSO per mL) were used for the vehicle in
each experiment. Staurosporine was purchased from Sigma and was used at 1
μM. For inhibitor experiments, 20
μM z-VAD-FMK, 200 nM wortmannin, 10
μM chloroquine, 10 nM bafilomycin A1,
or 10 μM necrostatin-1 (all Sigma) were added for 1
h prior to, and for the duration of the stimulation. The human TNF
neutralizing antibody (D1B4) was purchased from Cell Signaling, and was
added 2 h prior to SM treatment at 1 μg
mL−1.
hydroxamic acid) were purchased from Selleck Chemicals. Concentrations of
drugs used were based upon previously published whole blood
Cmax and
Cmin data for each drug (Amaravadi et al., 2015 (link); Flygare et al., 2012 (link); Zutshi et al., 1990 (link)) and were prepared in
dimethyl sulfoxide. Corresponding volumes of DMSO
(5μL DMSO per mL) were used for the vehicle in
each experiment. Staurosporine was purchased from Sigma and was used at 1
μM. For inhibitor experiments, 20
μM z-VAD-FMK, 200 nM wortmannin, 10
μM chloroquine, 10 nM bafilomycin A1,
or 10 μM necrostatin-1 (all Sigma) were added for 1
h prior to, and for the duration of the stimulation. The human TNF
neutralizing antibody (D1B4) was purchased from Cell Signaling, and was
added 2 h prior to SM treatment at 1 μg
mL−1.