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. Author manuscript; available in PMC: 2015 Sep 12.
Published in final edited form as: Oncogene. 2014 Mar 31;34(11):1341–1353. doi: 10.1038/onc.2014.72

Figure 1.

Figure 1

Figure 1

Figure 1

Piperlongumine inhibits cytoplasmic-to-nuclear translocation of Stat3 assessed by confocal and high-throughput fluorescence microscopy (HTFM). A, B. MEF/GFP-Stat3α (A) or MEF/GFP-Stat3β (B) cells were pretreated with vehicle control (DMSO; column 2) or (0.1/0.3/1/3/10/30/100 μM) of PL (columns 3–8) for 1 hour and stimulated without (column 1) or with IL-6 + IL-6sR (150 ng/ml) for 30′ (columns 2–8). Plates were examined by confocal fluorescent microscopy using filters to detect GFP (row 1), DAPI (row 2) or both (merge; row 3). C. MEF-GFP-Stat3α (left) or MEF-GFP-Stat3β (right) cells, treated as above, fixed and examined by HTFM to determine the fluorescence intensity in the nucleus (FLIN). Percent of maximum change (delta, D) in FLIN was plotted as a function of Log [M] PL. Data shown (mean ± SD) representative of ≥ 2 experiments. Best-fit curves were determined and IC50 values determined (Table 1) using 4-Parameter Logistic Model/Dose Response using GraphPad