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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: Emilin-2 is a component of bone marrow extracellular matrix regulating mesenchymal stem cell differentiation and hematopoietic progenitors

Fig. 2

Emilin-2 expression in ST2 cells during in vitro adipogenic differentiation. A mRNA levels of adipogenic markers in ST2 cells under non-differentiating conditions (UT t3) or after 3–7 days of treatment with adipogenic stimuli (Adipo t3 and t7), as determined by RT-qPCR. mRNA levels are shown as fold change compared to UT t3 condition (n = 3–9; *P < 0.05;**P < 0.01; ***P < 0.001). B RT-qPCR analysis of mRNA levels of Emilin-2 in ST2 cells under non-differentiating conditions (UT t0 and t3) and after 3 and 7 days of treatment with adipogenic stimuli (Adipo t3 and t7). mRNA levels are shown as fold change compared to UT t0 condition (n = 3–9; *P < 0.05; ***P < 0.001). C Western blot analysis for Emilin-2, AdipoQ and FABP4 proteins in ST2 cells under non-differentiating condition (UT t0 and t3) and after 3 days of adipogenic stimuli (Adipo t3). Three independent samples are shown for each condition. β-actin was used as a protein loading control. The graph on the right show densitometric quantification for Emilin-2, as determined by two independent experiments. Protein levels are shown as fold change compared to UT t3 condition (n = 3–6; *P < 0.05;**P < 0.01) D Immunofluorescence for Emilin-2 (green) in ST2 cells at 0, 3 and 7 days culture in untreated conditions (UT) or in the presence of adipogenic stimuli (Adipo). Nuclei were stained with Hoechst (blue). Scale bar, 100 µm. E, F RT-qPCR analysis of the mRNA levels for AdipoQ, FABP4 (E) and Emilin-2 (F) in ST2 cells cultured for 3 days before adipogenic treatment (day 3, UT t0) and then treated for 3 and 7 days with adipogenic stimuli (day 3, Adipo t3 and t7). mRNA levels are shown as fold change compared to day 3, UT t0 condition(n = 3; ***P < 0.001). G Relative mRNA levels of AdipoQ and FABP4 in ST2 cells cultured under non-differentiating conditions (UT t3) or after 3 days of adipogenic stimulus in cells pre-treated (Adipo + E2 t3) or not (Adipo t3) with purified Emilin-2 protein. mRNA levels are shown as fold change compared to UT t3 condition (n = 3; **P < 0.01; ***P < 0.001). See also Additional file 1: Figs. S1 and S2

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