Figure 2

Observation and adipogenic-specific RNAs quantification of adipocytes at 0, 7, 14 and 21 days of differentiation. (A) Images obtained by optical microscopy on human MSC-derived adipocytes (hMSC-Adi) after 0, 7, 14, and 21 days of differentiation. Scale bars: 100 μm. (B) RT-qPCR analysis of PPARγ, Leptin, CEBPα and CEBPδ in 0, 7, 14, and 21 days-differentiated adipocytes. Transcript levels of GAPDH were used for sample normalization. Data was obtained from 4 independent experiments (Bars: median ± interquartile space). Differences were considered significant at p < 0.05 (*p < 0.05, **p < 0.01). (n. d. not detected). (C) RT-qPCR of miR-138, miR-30c, miR-125a, miR-125b, miR-31 in 0 and 14 days-differentiated adipocytes. Transcript levels of RNU6P were used for sample normalization. (D) RT-qPCR analysis of PPARγ in 14 days-differentiated adipocytes (hAdi D14) transfected with siPPARγ or with non-target control siRNA (siCTRL). Transcript levels of 36B4 were used for sample normalization. (E) RT-qPCR analysis of PPARγ in 14 days-differentiated osteoblasts incubated with conditioned medium from 0 (ctrl) or 14 days-differentiated adipocytes transfected (hAdi D14 siPPARγ-CM) or not (hAdi D14-CM). Transcript levels of 36B4 were used for sample normalization.