Triptolide and Hypoxia Signaling Modulation

MIA PaCa-2 cells (ATCC) were cultured in DMEM medium (Hyclone) supplemented with 10% FBS and 1% penicillin streptomycin. S2-VP10 cells were cultured in RPMI 1640 medium (Hyclone) with 10% FBS and 1% penicillin streptomycin. Triptolide was used at a concentration of 50 nM in all in vitro experiments based on the response of this compound from our previous studies^{18 (link), 20 (link), 21 (link)} (unless otherwise mentioned). The hypoxia mimetic cobalt chloride (CoCl₂) (Sigma) was used at a concentration of 200 uM for HIF-1α reporter assays. The proteasome inhibitor MG-132 (Sigma) was used at a concentration of 10 uM for HIF-1α reporter assays. BAY87-2243, a hypoxia inhibitor was used in indicated concentration. Minnelide, the pro drug of triptolide, was used in vivo at indicated concentrations.