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Fig. 5 | BMC Medicine

Fig. 5

From: Three tyrosine kinase inhibitors cause cardiotoxicity by inducing endoplasmic reticulum stress and inflammation in cardiomyocytes

Fig. 5

The 3 TKIs induce different levels of lipid peroxidation, ROS, calcium defects, and TNNT2 loss in NRCMs. To measure oxidative stress and cardiotoxic effects induced by TKIs, NRCMs were treated with 10 µM afatinib, 10 µM sorafenib, or 5.62 µM ponatinib and stained with various dyes before quantification by flow cytometry or imaging. A-B Percent of ROS-high cells quantified by flow cytometry and staining with H2DCFDA after different TKI treatments for 3 or 24 h. C–D Histogram of the ratios between green (oxidized) and red (non-oxidized) fluorescence intensity of C11-bodipy581/591 in NRCMs under different treatments. Percentages by the condition names were the fraction of cells in the specified gate for each treatment. E Representative fluorescence images of live NRCMs pre-loaded with Calbryte™ 520 AM and treated with the indicated drug from 0 to 3 h. F Integrated density of green fluorescence in E. G Representative immunofluorescence images of TNNT2 (green) and nuclei (blue) in NRCMs treated with TKIs for 24 h. H Integrated density of green fluorescence in G. Data were presented as mean ± SEM (n = 3) and analyzed using ANOVA analysis. *p < 0.05, **p < 0.01, ***p < 0.001 versus the DMSO vehicle control group. Scale bar: 200 µm

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