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. 2017 Jan 27;7:41525. doi: 10.1038/srep41525

Figure 5. DTT prevents oridonin-induced activation of HSF1 and inhibitory effects in Ph+ leukemia cells.

Figure 5

(a) Cells were pre-incubated with DTT (1 mM) for 2 hours and then treated with oridonin (K562: 20 μM for 2 hours; KU812 and SUP-B15: 10 μM for 4 hours) before subjected to immunoblotting. (b) Immunostaining showing the absence of HSF1 granules (red) in nucleus (blue) of oridonin-treated cells in the presence of DTT. (c) DTT prevents oridonin-induced HSF1 trimerization. K562 cells were treated with oridonin for 2 hours in the absence or presence of DTT. Lysates were cross-linked with EGS before subjected to immunoblotting. (d) DTT inhibits oridonin-induced transactivation of hsp70 and ubc. Cells were treated with oridonin for 24 hours with or without DTT and subjected to qPCR analysis. (e) DTT abrogates the inhibition of cell growth by oridonin as assessed by MTT assays. (f) Oridonin-induced cell apoptosis in Ph+ leukemia cells was largely antagonized by DTT as determined by Annexin V/PI staining and flow cytometry.