sureprint g3 human 8 × 60 k oligo-microarray chip  (Agilent technologies)

Journal: Stem Cells International

Article Title: Romidepsin Promotes Osteogenic and Adipocytic Differentiation of Human Mesenchymal Stem Cells through Inhibition of Histondeacetylase Activity

doi: 10.1155/2018/2379546

Figure Lengend Snippet: Microarray gene expression profiling of adipocyte-differentiated human bone marrow stromal stem cells (hMSCs) following romidepsin treatment. (a) Heat map and unsupervised hierarchical clustering were performed on differentially expressed genes in romidepsin-treated hMSCs compared to those in vehicle-treated control hMSCs on day 7 after adipocyte differentiation. (b) Pie chart illustrating the distribution of the top 15 enriched pathway categories in the differentially expressed genes identified in romidepsin-treated hMSCs. (c) Validation of a selected group of upregulated genes identified by microarray analysis using qRT-PCR. Gene expression was normalized to β -actin. Data are presented as mean fold changes ± SEM compared with vehicle-treated controls; n = 6 from two independent experiments, ∗ P < 0.05 and ∗∗∗ P < 0.0005 comparing romidepsin-treated and control cells. (d) Quantification of Nile red staining for mature adipocytes in romidepsin-treated hMSCs and in the absence or presence of a FAK inhibitor (5 μ M). Data are presented as mean ± SEM, n = 6, ∗∗ P < 0.005, and ∗∗∗ P < 0.0005.

Article Snippet: The total RNA (150 ng) was labeled using the low input Quick Amp Labeling Kit (Agilent Technologies, Santa Clara, CA, USA), then hybridized to the Agilent SurePrint G3 Human GE 8 × 60 k microarray chip (Agilent Technologies).

Techniques: Microarray, Expressing, Quantitative RT-PCR, Staining

Journal: Stem Cells International

Article Title: Romidepsin Promotes Osteogenic and Adipocytic Differentiation of Human Mesenchymal Stem Cells through Inhibition of Histondeacetylase Activity

doi: 10.1155/2018/2379546

Figure Lengend Snippet: Microarray gene expression profiling of osteoblast-differentiated human bone marrow stromal stem cells (hMSCs) following romidepsin treatment. (a) Heat map and unsupervised hierarchical clustering were performed on differentially expressed genes induced by romidepsin compared to those of vehicle-treated control hMSCs at day 10 after osteoblastic differentiation; (b) Pie chart illustrating the distribution of the top 15 enriched pathway categories in the differentially expressed genes identified in romidepsin-treated hMSCs; (c) validation of the selected gene panel during osteoblastic differentiation using qRT-PCR. Gene expression was normalized to β -actin; (d) quantification of ALP activity in hMSCs induced to osteoblastic differentiation for 10 days after treatment with romidepsin in the absence or presence of a FAK inhibitor (PF-573228, 5 μ M) or TGF β inhibitor (SB505124, 5 μ M). Data are presented as mean ± SEM, n = 8; ∗ P < 0.05, ∗∗ P < 0.005, and ∗∗∗ P < 0.0005.

Article Snippet: The total RNA (150 ng) was labeled using the low input Quick Amp Labeling Kit (Agilent Technologies, Santa Clara, CA, USA), then hybridized to the Agilent SurePrint G3 Human GE 8 × 60 k microarray chip (Agilent Technologies).

Techniques: Microarray, Expressing, Quantitative RT-PCR, Activity Assay