Fig. 5

Enhanced osteogenic and adipogenic differentiation ability in obese ASCs treated with MLN or PD accompanied by downregulation of the transcription factors KLF4 and KLF6. a–d Obese ASCs, pretreated for 72 h with MLN or PD, were induced into the osteogenic differentiation. The percentage of differentiated ASCs was evaluated by alizarin red S staining. a, b The results are presented as median ± min/max whiskers (red dashed line indicates median value of obese ASCs) in visceral and subcutaneous ASCs (n = 300 cells for each condition, pooled from three experiments). Example images are shown in c and d, respectively. Scale bars 30 μm. e Gene analyses of visceral ASCs with indicated conditions for RUNX2 and PTCH1, two genes involved in osteogenesis. The results are from three experiments and presented as mean ± SEM. f Gene levels of KLF4 and KLF6 after osteogenic differentiation in visceral obese ASCs. The results are from three experiments, presented as mean ± SEM. h Gene Levels of ADIPOQ, LEPTIN, and PPARγ after adipogenic differentiation are shown for obese visceral ASCs. The results are from three experiments, presented as mean ± SEM. g Quantification of cells showing lipid vacuoles after 14 days of adipogenic differentiation. The results are presented as median ± min/max whiskers in visceral ASCs (n = 150 cells for each condition, pooled from three experiments), and the red dashed line illustrates the median value of obese ASCs. Box and whisker plots were used to show the median and the minimal to maximal range of the values in a, b, and g. Unpaired Mann-Whitney U test for a, b, and g. Student’s t test for e, f, and h. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001