Research Article
5-Azacytidine and Resveratrol Enhance Chondrogenic Differentiation of Metabolic Syndrome-Derived Mesenchymal Stem Cells by Modulating Autophagy
Figure 6
Investigation of autophagy during early chondrogenesis. Prior to experiments, cells were pretreated with AZA/RES for 24 hours, then the experimental medium was replaced by osteogenic differentiation medium. Cells were cultured in that medium for five days and after were subjected to further analysis. Nuclei of cells were stained with DAPI. Mitochondria were stained with MitoRed dye while LAMP-2 was visualized using immunofluorescence. Mitochondrial net and LAMP-2 were visualized in cells using confocal microscopy (a). Expression of genes related to autophagy was investigated with RT-PCR. Expression of Beclin-3 (b) and LC3 (c) was diminished in EMS EXP; however, LAMP-2 (d) and p62 (e) mRNA levels were upregulated in that group. Similarly, mTOR expression was diminished (f). Results expressed as . Statistical significance indicated as an asterisk () when comparing the result to ASCEMS and as a number sign (#) when comparing to ASCCTRL. ; ### and .
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