Research Article

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

Figure 4

IL21R gene is modulated during adipogenesis, which is affected by gene silencing. (a, b) Relative quantification of mRNA IL21R and STAT3 during adipogenesis in ADSCs, measured using qRT-PCR. The data were performed in biological and technical triplicate (TAL01, 09, and 32), normalized by POLR2A. Mean with SEM, two-way ANOVA with multiple comparisons: , , ns: not significant. (c–f) siRNA-treated cells were induced to adipogenesis for ten days. The cells were then incubated with Nile Red (lipophilic fluorescent dye) and analyzed using the Operetta High-Content Imaging System (TAL23, 27, and 32). (c) Representative fluorescence images of ADSCs silencing IL21R and scramble after ten days of adipogenesis and stained with Nile Red (green staining) and DAPI (blue staining), representative images of the donor TAL23. Scale bar: 200 μm. (d, f) Nile Red fluorescence quantification of ADSCs silenced for IL21R and induced to adipogenic differentiation for ten days. (d) Percentage of Nile Red positive cells, (e) mean fluorescence intensity per cell, and (f) the number of lipid vesicles per cell. Mean with SEM; Student’s unpaired -test analysis: , ns: not significant.
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