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Figure 5: Confocal immunofluorescence imaging of the DhRho 123 detectable ROS/RNS products in MSCs challenged with LPS. (a1) Projection of oxidized form of DhRho 123 (Rho 123) (green channel) in control MSCs. (a2) Overlay of projection of Rho 123 shown in (a1) and a respective DIC image. (b1) Projection of Rho 123 (green channel) in LPS-challenged MSCs. (b2) Overlay of projection of Rho 123 shown in (b1) and a respective DIC image. A dramatic increase in Rho 123 fluorescence occurred in the LPS-challenged MSCs. (c1) Projection of Rho 123 (green channel) in LPS-challenged MSCs and treated with LNIL, an iNOS inhibitor. (c2) Overlay of projection of Rho 123 shown in (c1) and a respective DIC image. Suppression of Rho 123 fluorescence occurred in the LPS-challenged MSCs. Bright green fluorescence of the ROS-activated Rho 123 shown in mitochondria is shown with red arrows. Diffused green fluorescence of the ROS/RNS-activated Rho 123 in the cytoplasm is shown with white arrows in (b). The confocal images were taken with pinhole setup to obtain 0.5 μm Z-sections. The experimental conditions were the same as indicated in Figure 2.