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HBO Pretreatment Activates the AKT/HSF-1 Pathway via <t>ROS</t> Elevation. ROS were labeled by mitochondrial-targeted <t>fluorescent</t> probe Mito-SOX during HBO exposure. Mitochondrial-targeted ROS scavenger Mito-Tempo and whole-cellular ROS scavenger NAC were applied 1 h before HBO. (a) Fluorescence staining of mitochondrial ROS (Red), nuclei were marked with DAPI (Blue) (scale bar: 20 μm); (b) HSF-1 nuclear translocation. ROS: reactive oxygen species. n = 3, * P < 0.05, ** P < 0.01.Western blot was used to detect the activation of signaling molecules. Specific inhibitors or ROS scavengers were applied before HBO exposure. (c–f) Phosphorylation levels of Akt, ERK1/2, p38 MAPK and MEK1/2 after HBO exposure; (g–h) HSF-1 nuclear translocation after inhibition of ROS or signaling molecules. (i) AKT phosphorylation after ROS elimination. n = 3, * P < 0.05, ** P < 0.01.
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HBO Pretreatment Activates the AKT/HSF-1 Pathway via ROS Elevation. ROS were labeled by mitochondrial-targeted fluorescent probe Mito-SOX during HBO exposure. Mitochondrial-targeted ROS scavenger Mito-Tempo and whole-cellular ROS scavenger NAC were applied 1 h before HBO. (a) Fluorescence staining of mitochondrial ROS (Red), nuclei were marked with DAPI (Blue) (scale bar: 20 μm); (b) HSF-1 nuclear translocation. ROS: reactive oxygen species. n = 3, * P < 0.05, ** P < 0.01.Western blot was used to detect the activation of signaling molecules. Specific inhibitors or ROS scavengers were applied before HBO exposure. (c–f) Phosphorylation levels of Akt, ERK1/2, p38 MAPK and MEK1/2 after HBO exposure; (g–h) HSF-1 nuclear translocation after inhibition of ROS or signaling molecules. (i) AKT phosphorylation after ROS elimination. n = 3, * P < 0.05, ** P < 0.01.

Journal: Frontiers in Molecular Biosciences

Article Title: Hyperbaric oxygen pretreatment on endothelial cell injury via heat shock factor 1 in decompression sickness

doi: 10.3389/fmolb.2025.1617318

Figure Lengend Snippet: HBO Pretreatment Activates the AKT/HSF-1 Pathway via ROS Elevation. ROS were labeled by mitochondrial-targeted fluorescent probe Mito-SOX during HBO exposure. Mitochondrial-targeted ROS scavenger Mito-Tempo and whole-cellular ROS scavenger NAC were applied 1 h before HBO. (a) Fluorescence staining of mitochondrial ROS (Red), nuclei were marked with DAPI (Blue) (scale bar: 20 μm); (b) HSF-1 nuclear translocation. ROS: reactive oxygen species. n = 3, * P < 0.05, ** P < 0.01.Western blot was used to detect the activation of signaling molecules. Specific inhibitors or ROS scavengers were applied before HBO exposure. (c–f) Phosphorylation levels of Akt, ERK1/2, p38 MAPK and MEK1/2 after HBO exposure; (g–h) HSF-1 nuclear translocation after inhibition of ROS or signaling molecules. (i) AKT phosphorylation after ROS elimination. n = 3, * P < 0.05, ** P < 0.01.

Article Snippet: Levels of intracellular ROS were measured using the mitochondria-targeted ROS fluorescent probe Mito-Sox (Thermo Fisher Scientific).

Techniques: Labeling, Fluorescence, Staining, Translocation Assay, Western Blot, Activation Assay, Phospho-proteomics, Inhibition