Fig. 4

NOX4 localizes at mitochondria and contributes to the increase in ROS production after sublethal heat stress. A RT-qPCR analysis of NOX1–5 and DUOX1–2 mRNAs expression after sublethal heat stress. B Western blot of NOX4 expression in total cell lysates after sublethal heat stress. C Colocalization of mitochondria and NOX4. MitoTracker™ red and FITC- conjugated secondary antibody was used for NOX4. Nuclei were counterstained with Hoechst 33,342. Yellow displays colocalization. D Western blot of NOX4 expression in mitochondria after sublethal heat stress. E Protein levels of NOX4 in whole cell extracts. F Protein levels of NOX4 in mitochondria. G DCFH-DA and H MitoSOX™ fluorescence as measured using flow cytometry at 6 h after sublethal heat stress.I and J Apoptosis was quantified using Annexin V-FITC/PI staining and flow cytometry at 24 h after sublethal heat stress. Values are the mean ± SE. *p < 0.05;***p < 0.001; ns no significance