Journal: Scientific Reports

Article Title: Mechanotransductive stabilization of HIF-1α is inhibited by mitochondrial antioxidant therapy in the setting of pulmonary overcirculation

doi: 10.1038/s41598-025-99062-0

Figure Lengend Snippet: Primary PAECs and snap frozen peripheral lung tissues from shunt and control animals were compared. ( A ) Shunt PAECs exhibit higher mitochondrial ROS production based on fluorescence intensity with MitoSOX staining. Results were analyzed as control and shunt pairs, with each shunt normalized to paired control PAEC fluorescence level. N = 3 shunt and 3 control cell lines. Scale bars = 50 μM ( B ) Shunt PAECs have significantly higher levels of HIF-1α protein by western blot. N = 4 shunt and N = 5 control cell lines. ( C ) Peripheral lung tissue from shunt animals shows significantly higher levels of superoxide content by EPR analysis ( D ) Peripheral lung tissue from shunt animals also demonstrates significantly higher levels of HIF-1α protein. N = 5 shunt and N = 5 control animals for tissue studies. Western blots HIF-1α levels are normalized to β-actin as an internal loading control and expressed as band intensity ratios. All p -values calculated by unpaired, 2-tailed t-test with p < 0.05 considered significant. For bar graphs * denotes p -value ≤ 0.05, ** denotes p -value ≤ 0.01, *** denotes p -value ≤ 0.001 and **** denotes p -value ≤ 0.0001.

Article Snippet: Fig. 6 Primary PAEC cell lines, serum and peripheral lung tissue derived from treated MitoQ shunt and untreated shunt animals were evaluated for NO levels and production. ( A ) MitoQ shunt PAECs exhibit increased NO production compared to untreated shunt cells by DAF FM fluorescence intensity.

Techniques: Control, Fluorescence, Staining, Western Blot

Journal: Scientific Reports

Article Title: Mechanotransductive stabilization of HIF-1α is inhibited by mitochondrial antioxidant therapy in the setting of pulmonary overcirculation

doi: 10.1038/s41598-025-99062-0

Figure Lengend Snippet: Shunt animals were treated daily with oral MitoQ. Primary PAEC cell lines were derived, and peripheral lung tissues collected and snap frozen from treated (note that all tissues and PAECs derived from these treated animals are labelled “MitoQ Shunt”) and untreated shunt animals for analysis. ( A ) MitoQ shunt PAECs exhibit lower mitochondrial ROS production than shunt PAECs based on fluorescence intensity with MitoSOX staining. Results were analyzed as control and shunt pairs, with each shunt normalized to paired control PAEC fluorescence level. N = 3 shunt and N = 3 control cell lines. ( B ) MitoQ shunt PAECs (N = 4) have significantly lower HIF-1α protein levels than shunt cells (N = 5) by Western Blot. ( C ) Peripheral lung tissue from MitoQ shunt animals (N = 4) demonstrates lower superoxide levels than shunt animals (N = 4) and no different from lung tissue from physiologically normal control animals (N = 4) by EPR with representative waves included for each group. Analysis was performed by 2-way ANOVA with α of .05 as the significance threshold. ( D ) Peripheral lung tissue from MitoQ shunt animals (N = 4) has significantly lower levels of HIF-1α protein than untreated shunts. Western blots HIF-1α levels are normalized to β-actin as an internal loading control and expressed as ratios of band intensity. All p -values calculated by unpaired, 2-tailed t-test with p < 0.05 considered significant.

Article Snippet: Fig. 6 Primary PAEC cell lines, serum and peripheral lung tissue derived from treated MitoQ shunt and untreated shunt animals were evaluated for NO levels and production. ( A ) MitoQ shunt PAECs exhibit increased NO production compared to untreated shunt cells by DAF FM fluorescence intensity.

Techniques: Derivative Assay, Fluorescence, Staining, Control, Western Blot

Journal: Scientific Reports

Article Title: Mechanotransductive stabilization of HIF-1α is inhibited by mitochondrial antioxidant therapy in the setting of pulmonary overcirculation

doi: 10.1038/s41598-025-99062-0

Figure Lengend Snippet: Primary PAEC cell lines, serum and peripheral lung tissue derived from treated MitoQ shunt and untreated shunt animals were evaluated for NO levels and production. ( A ) MitoQ shunt PAECs exhibit increased NO production compared to untreated shunt cells by DAF FM fluorescence intensity. Results analyzed as control and shunt pairs, with each shunt normalized to paired control PAEC fluorescence level. N = 3 shunt and N = 3 control cell lines. ( B ) Culture media from MitoQ shunt PAECs contains higher levels of NO x than media from untreated shunt cells. (N = 3 MitoQ shunt and N = shunt cell lines) ( C ) Plasma samples and ( D ) peripheral lung tissue from MitoQ shunt animals have significantly higher NO x levels than corresponding samples from untreated shunt animals, though not as high as physiologically normal control animals. (N = 3 MitoQ shunt, shunt, and normal control animals per group performed on samples in triplicate). p -values for A&B calculated by unpaired, 2-tailed t-test with p < 0.05 considered significant. Analysis for C&D was performed by 2-way ANOVA with α of .05 as the significance threshold.

Article Snippet: Fig. 6 Primary PAEC cell lines, serum and peripheral lung tissue derived from treated MitoQ shunt and untreated shunt animals were evaluated for NO levels and production. ( A ) MitoQ shunt PAECs exhibit increased NO production compared to untreated shunt cells by DAF FM fluorescence intensity.

Techniques: Derivative Assay, Fluorescence, Control, Clinical Proteomics