Fig. 3.

RECK differentially controls adipogenesis and osteogenesis in hMSCs. hMSCs were cultivated for 3 and 14 days in the absence (con) and presence of adipogenic (adipo) and osteogenic (osteo) differentiation medium. a Relative transcriptional levels of RECK were quantified using qRT-PCR and normalized to GAPDH mRNA. b RECK present in cell extracts was determined by Western blot analysis applying equal amounts of total protein into each lane. Results of densitometric quantification are given in densitometric units (DU) with the amounts of protein present in control cells set as 100 %. hMSCs transfected with control siRNA (NC) or siRNA targeting RECK (KD) were incubated in adipogenic or osteogenic differentiation medium. c Relative mRNA expression of the differentiation markers PPARγ and ALP was quantified by qRT-PCR analysis after 10 days of differentiation. Values were normalized to GAPDH mRNA. Cell staining with Oil Red O for adipogenic and Alizarin Red S for osteogenic differentiation after 14 days of incubation in differentiation medium is shown by representative microscopic images of stained cellular monolayers (d) and stain recovery after extraction from the cells (e). Scale bars indicate 1 mm. Data shown represent the mean ± SD of triplicate measurements (n = 3). *P < 0.05; **P < 0.01; ***P < 0.001