Hypoxia Rapidly Induces the Expression of Cardiomyogenic Factors in Human Adipose-Derived Adherent Stromal Cells

Figures

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  Figure 1. Experimental scheme of this study (A) and surface protein expression on adipose-derived adherent stromal cells (ADASs) and representative adipose-derived stem cells (ASCs), as measured by flow cytometry (B). Black histograms show the expression of isotype controls and blue and red histograms indicate the expression of target proteins on ASCs and ADASs, respectively. The data are representative of two independent experiments.

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  Figure 2. Time-dependent differential regulation of differentiation-related genes of cell lineages in ADASs under normoxic or hypoxic conditions as determined by qRT-PCR. The data are representative of three independent experiments. N, normoxia; H6, hypoxia for 6 h; H12, hypoxia for 12 h; H24, hypoxia for 24 h. Statistical significance between the normoxic and hypoxic groups is indicated in Table S1.

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  Figure 3. Time-dependent differential regulation of cardiomyogenic proteins in ASCs and ADASs under normoxic and hypoxic conditions as determined by immunoblot analysis. Band intensity was measured as area density and analyzed in ImageJ. Relative intensity levels indicate protein levels normalized to β-actin levels. The data are representative of two independent experiments. N, normoxia; H6, hypoxia for 6 h; H12, hypoxia for 12 h; H24, hypoxia for 24 h. Statistical significance between the normoxic and hypoxic groups is indicated in Table S2. Western images are shown in Supplementary figures.

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  Figure 4. Expression of cardiomyogenic proteins, GATA binding protein-4 (GATA-4) and troponin T in ASCs and ADASs in response to hypoxia (24 h) as determined by immunocytochemical staining. Nuclei were stained with DAPI. Scale bar = 100 μm.

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  Figure 5. Changes in cardiomyogenic factors in dimethyloxaloylglycine (DMOG)-treated ASCs and ADASs as determined by qRT-PCR (A) and immunoblot analysis (B). All values of qRT-PCR are shown as the normalized target gene expression level relative to GAPDH transcript levels. p values indicated as * p < 0.05 and ** p < 0.01 between the control group and DMOG-treated group. The data are representative of two independent experiments. Immunoblot images are shown in the supplementary figure.

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  Figure 6. Secretome analysis in a conditioned medium (CM) of ASCs and representative ADASs under normoxic or hypoxic conditions. Statistical significance between hypoxic ASCs and hypoxic ADAS and between normoxic ADASs and hypoxic ADASs was determined by ANOVA using SPSS, with p values indicated as * p < 0.05. N, normoxia; H24 hypoxia for 24 h.