Molecular response to PTL and temsirolimus. AML samples were treated with parthenolide (PTL), alone or in combination with 2.5 μg/mL of temsirolimus (TEM). (A) Confocal micrograph for TEX cells after treatment for 6 hours with either 2.5 μg/mL TEM, 5μM PTL, either alone or in combination. Cells were stained with anti-Nrf2 antibody (green) and nuclear dye DAPI (4,6 diamidino-2-phenylindole, blue). (B) qPCR analyses of heme oxygenase I in CD34⁺ AML specimens after 6 hours of treatment with PTL, TEM, or PTL + TEM (P + T). Data are shown relative to untreated controls for AML1-AML7. Fold changes are represented on a log-10 scale (log₁₀RQ). (C) Immunoblot analyses for the indicated treatments at the 6-hour timepoint. Blots were probed with antibodies against either heme oxygenase 1 (HO-1) or β-actin. (D) qPCR analyses of GCLM (left panel; AML1, AML2, AML4, AML5, AML6) and NQO1 (right panel; AML1-AML7) in CD34⁺ AML specimens after 6 hours of treatment with PTL, TEM, or PTL + TEM (P + T). Data are shown relative to untreated controls. Fold changes are represented on a log-10 scale (log₁₀RQ).