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. 2018 May 5;12(6):869–882. doi: 10.1002/1878-0261.12199

Figure 2.

Figure 2

Effect of BRAFV600E on ROS production in PEITC‐treated NIH 3T3 wt and NIH 3T3 BRAFV600E cells. (A) A total of 60 000 cells were seeded in 8‐well Nunc Lab‐Tek chambers and after 20 h treated with PEITC (20 μm) for 30 min and afterward stained with MitoTracker Red CM‐H2XRos. Data are presented as mean value ± standard error of the mean (SEM) of 100 measurements per sample and three independent experiments. Size bar: 50 μm (B) FACS analysis after CM‐H2DCFDA staining of NIH 3T3 wt and NIH 3T3 BRAFV600E cells and PEITC treatment. A total of 500 000 cells were seeded in six‐well plates, and after 30 min of treatment with different concentrations of PEITC, they were stained with CM‐H2DCFDA and subsequently analyzed by FACS. Data are presented as mean value ± standard error of the mean (SEM) of three experiments. (C) A total of 500 000 cells were seeded in six‐well plates and treated overnight with 20 μm PEITC before performing FACS analysis after annexin V staining. Data are presented as mean value ± standard error of the mean (SEM) of three experiments. *P < 0.1, **P < 0.01, ***P < 0.001.