Figure 1.

Compromise in endothelial CSE by siRNA silencing enhances oxidative status in HUVEC. (A) CSE gene and protein expression were determined by RT-PCR and Western blot, respectively in CSE and control siRNA transfected HUVEC. Statistical significance was measured via Mann–Whitney test. (B) Intracellular H₂S availability was measured by fluorescence microscopy using the specific fluorescence probe SF7-AM. Statistical significance was measured via unpaired T test. (C) Intracellular reactive oxygen species were detected by fluorescence microscopy using the fluorescent probe DCFDA. Statistical significance was measured via Mann–Whitney test. (D) Mitochondrial superoxide was measured by fluorescent microscopy using MitoSOX Red probe. Statistical significance was measured via unpaired T test. A shows a representative Western blot from n = 1 experiment. As indicated in the materials and methods section, Western blot membranes were cropped for consise display. Microscopy images were taken using the 20X objective lens ans indicated in the materials and methods section. Values are expressed as means ± SEM and each dot represents a biological replicate examined in three independent experiments. n = 3–6 experimental replicates. *P < 0.05, **P < 0.01, ***P < 0.001 ****P < 0.0001 vs siCTL. Figure was created using GraphPad Prism 8.1 (GraphPad Software, La Jolla, CA; https://www.graphpad.com/scientific-software/prism/).