Fig. 5

26S proteasome functional status under hypoxia determines the integrity of mitochondria dynamics in human BM-MSCs. Human BM-MSCs were incubated with MG132 inhibitor(10 µm), and two different hypoxia percentages: 2.5% (moderate hypoxia) and 0.5% (sever hypoxia) for 24 h. TMRE Mitochondrial Membrane Potential Assay was performed. a, b The TMRE fluorescence intensity readings and imaging in human BM-MSCs treated with MG132 inhibitor is enormously low with similar results found in severe hypoxia compared to normoxia and moderate hypoxia. On the contrary, moderate hypoxia-treated human BM-MSCs showed sufficient TMRE intensity readings close to those found in normoxia. n = 6; b live fluorescent images showed a weak signal of TMRE stain in human BM-MSCs after using the 26S MG132 and severe hypoxia weighed against normoxia and moderate hypoxia where the signal was strong and bright. b Mitochondrial complexes activity assays of human BM-MSCs after being subjected to experimental treatments. The activity of most complexes was negatively affected in human BM-MSCs treated with MG132 or severe hypoxia with the most discernable decrease in complexes IV and V, while the activity of mitochondrial complexes continued in normal mode in human BM-MSCs treated with moderate hypoxia similar to normoxia state; n = 3. *p < 0.05 compared to normoxia group; #p < 0.05 compared to moderate hypoxia. This experiment was repeated 3 times