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Additional Supporting Information may be found in the online version of this article.

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sc-12-0114_sm_SupplFigure1.tif242KFigure S1: Immunostaining for Nanog in nBM-MSC (green). Cell nuclei were counterstained with Hoechst 33342 (blue).
sc-12-0114_sm_SupplFigure2.tif18KFigure S2: Nanog alters the transcription level of p16INK4a regulators. Quantitative RT-PCR for (A) MEOX2 and (B) EZH2. Expression level of the indicated transcripts was normalized to GAPDH. The relative expression level was defined as the ratio of the normalized expression level of each gene in BM-MSCs to that in control nBM-MSCs. Real-time RT-PCR experiment was repeated with RNA from three independent experiments. (*) p< 0.05, n=3.
sc-12-0114_sm_SupplFigure3.tif335KFigure S3: Karyotype analysis based on DAPI banding (nBM-MSC) or Giemsa banding (aBMMSC) of Nanog expressing BM-MSC showed 54 diploid chromosomes as expected for normal ovine cells.
sc-12-0114_sm_SupplFigure4.tif44KFigure S4: Nanog increased transcription of TGF-β1 in nBM-MSC and aBM-MSC. Realtime qRT-PCR for TGF-β1. GAPDH served as a loading control. Results from 3 independent experiments were plotted as average±SE. (*) p<0.05, n=3.
sc-12-0114_sm_SupplFigure5.tif216KFigure S5: Nanog did not affect the amount of total Smad2. (A) Western blotting for total Smad2 in nBM-MSC and aBM-MSC; GAPDH served as loading control. (B) The band intensity of total Smad2 was determined using Image J software and normalized to GAPDH. Results from 3 independent experiments were plotted as average ± SE.
sc-12-0114_sm_SupplFigure6.tif199KFigure S6: TGF-β-neutralizing antibody inhibited nBM.N-CM-induced hydrogel contraction and Smad2 phosphorylation. (A) Exogenous TGF-β1 (2ng/ml) was neutralized by different concentrations of anti-TGF-β neutralizing antibody (R&D systems, Minneapolis, MN) as indicated and the level of p-Smad2 in control nBM-MSCs was measured by WB. (B) Gel compaction profile and (C) p-Smad2 level of control nBM-MSCs in the presence of nBM.C-CM or nBM.N-CM with or without TGF-β neutralizing antibody (5μg/mL) as indicated. (*) p<0.05 between the indicated samples and corresponding control at the same time point; (#) p<0.05 between the indicated sample and nBM.N-CM treated sample at the same time point (n=3).
sc-12-0114_sm_SupplFigure7.tif20KFigure S7: Exogenous TGF-β1 pathway increased Nanog-induced hydrogel compaction. Gel compaction profiles of (A) control BM-MSCs and (B) Nanog+ BM-MSCs with or without TGF-β1 (2 ng/ml). The TGF-β receptor inhibitor SB431542 (10 μM) was added in some samples as indicated.
sc-12-0114_sm_SupplFigure8.tif27KFigure S8: Differential effects of Oct4 on proliferation and contractility of nBM-MSC and aBM-MSC. (A) Number of colonies larger than 2mm in diameter from the clonogenic assay. (B) Doubling time. (C) Vascular reactivity (kPa) of nBM-MSC and aBM-MSC-derived TEVs in response to Endothelin-1 (20 nM), U46619 (10-6 M) or KCl (118 mM). All values are the mean±SE of three independent experiments (n=3). The symbol (*) denotes p<0.05 between the indicated samples and their corresponding controls.
sc-12-0114_sm_SupplTable1.xls7847KTable S1: Genes expressed in at least one of four cells: aBM.C, aBM.N, nBM.C, or nBM.N. Fold changes of up- and down-regulated genes (FC ≥ 2, p<0.05) are highlighted red and green, respectively and p-values of less than 0.05 are highlighted yellow.
sc-12-0114_sm_SupplTable2.xls630KTable S2: Venn diagram depicting DEG in each of the three group comparisons. Fold changes of up- and down-regulated genes (FC ≥ 2, p<0.05) are highlighted red and green, respectively and p-values of less than 0.05 are highlighted yellow. A: All DEG in at least one of the three group comparisons B: Common DEG between nBM.C vs. aBM.C and aBM.N vs. aBM.C groups C: Common DEG between nBM.C vs. aBM.C and nBM.N vs. nBM.C groups D: Common DEG among all three groups
sc-12-0114_sm_SupplTable3.xls56KTable S3: Estimated list of DEG promoters in nBM-MSC and aBM-MSC with Nanog binding sites.
sc-12-0114_sm_SupplTable4.xls99KTable S4: Gene ontology classification of DEG in each of the three group comparisons.
sc-12-0114_sm_SupplTable5.pdf43KTable S5: Primer sequences used for RT-PCR.

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