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. Author manuscript; available in PMC: 2012 Mar 13.
Published in final edited form as: Chem Biol. 2011 Nov 23;18(11):1355–1361. doi: 10.1016/j.chembiol.2011.09.008

Figure 1. Induction of NQO1, HO-1, and Hsp70 by Celastrol.

Figure 1

(A) Chemical structure of celastrol.

(B) Hepa 1c1c7 cells (104 per well) in 96-well plates were exposed to serial dilutions of the quinone methide triterpenoid celastrol. NQO1 activity was measured 48 hr later in cell lysates. Results are mean values (n = 8). The SD in each case was <10% of the value. CD, Concentration that doubles the specific activity of NQO1.

(C) Wild-type (WT) or HSF1-knockout (KO) MEFs (2 × 105 per well) in 6-well plates were exposed to vehicle (0.1% DMSO) or increasing concentrations of celastrol (CL) for 4 hr. Cells were lysed in RIPA buffer 20 hr later, aliquots from cell lysates were resolved by SDS/PAGE, transferred to immobilon-P, and probed with antibodies against Hsp70 (StressMarq Biosciences Inc.; 1:1000 dilution), HO-1 (Enzo Life Sciences; 1:1000 dilution), and NQO1 (1:3000 dilution; a kind gift from John D. Hayes, University of Dundee). The antibody against glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Sigma-Aldrich; 1:5000 dilution) was used as a loading control. The data are representative of two independent experiments.