Interleukin 21 Receptor Affects Adipogenesis of Human Adipose-Derived Stem/Stromal Cells

Figure 2

Overexpression of the IL21R gene in ADSCs. Cells were transfected with plasmid to overexpression of IL21R (IL21R), with a truncated plasmid (pcDNA) as a control. (a) Relative quantification of IL21R RNA normalized by GAPDH using qRT-PCR () (TAL22, 27, and 32). (b) Western blot analysis to verify IL21R protein levels () (TAL22, 27, 32, and 36). Protein levels of GAPDH were used as a control in the load, representative images of the donor TAL22. (c) Mitochondrial probe JC-10 measuring mitochondrial membrane potential () in ADSCs with overexpressed IL21R () (TAL23, 27, and 32). JC-10 fluorescence ratio at basal () and uncoupling states () was used to measure . DMSO was used as a control. (d, e) Rhodamine 123 assay to measure after overexpression of IL21R () (TAL23, 27, and 32). (d) Representative histogram with mitochondrial activity analysis. Cells without rhodamine (red curve) were used as a negative control. (e) Quantification of the relative fluorescence intensity means of rhodamine-stained cells. (f, g) Proliferation analysis using EdU incorporation in ADSCs with overexpressed IL21R () (TAL22, 28, and 32). (f) Representative histogram with proliferation analysis by EdU incorporation. Cells without EdU (red curve) were used as a negative control. (g) Percentage of EdU-positive cells. Mean with SEM; Student’s unpaired -test analysis: , ns: not significant.

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