1. A Tumor Necrosis Factor-α and Hypoxia-Induced Secretome Therapy for Myocardial Repair 
Kaviyanka Selvasandran, MS, Georges Makhoul, MD, MS, Prashant K. Jaiswal, PhD, Rishi Jurakhan, MS, Li Li, MS, Khalid Ridwan, MD, MS, Renzo Cecere, MD 
The Annals of Thoracic Surgery 
Volume 105, Issue 3, Pages (March 2018)
DOI: /j.athoracsur
Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

2. Fig 1
Immunofluorescence reveals the expression of tumor necrosis factor receptors (TNFR)-1 and -2 (green) on the plasma membranes of (A and D) unconditioned and (B, C, E, F) preconditioned rat bone marrow mesenchymal stem cells. 4,’6-diamidino-2-phenylindole (DAPI) stains, visualized in blue, represent the nuclei of the cells (original magnification ×20). (CM = conditioned medium; TNF = tumor necrosis factor.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

3. Fig 1
Immunofluorescence reveals the expression of tumor necrosis factor receptors (TNFR)-1 and -2 (green) on the plasma membranes of (A and D) unconditioned and (B, C, E, F) preconditioned rat bone marrow mesenchymal stem cells. 4,’6-diamidino-2-phenylindole (DAPI) stains, visualized in blue, represent the nuclei of the cells (original magnification ×20). (CM = conditioned medium; TNF = tumor necrosis factor.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

4. Fig 2
(a) Western blot analysis was conducted on rat bone marrow mesenchymal stem cells (rBM-MSC) that were initially treated with combinations of H9c2 cardiomyoblast–conditioned media (CM), normoxia, hypoxia, and tumor necrosis factor (TNF)-α (50 ng/mL). Protein expression was normalized to β-actin. Increased expression of (c) angiopoietin-2 (Ang-2), (d) fibroblast growth factor (FGF)-2, (f) vascular endothelial growth factor (VEGF)-1, and (g) transforming growth factor (TGF)-β in TNF-α + H9c2-CM + hypoxia–treated cells was indicated compared with H9c2-CM + hypoxia–treated cells. (b) Ang-1 expression is decreased, whereas (e) FGF-7 expression remains unchanged. Fold differences were normalized to the untreated rBM-MSC group. The error bars on the graphs represent the SD. Statistical significance is at p < 0.05. *p < 0.05, **p < 0.01, ***p < 0.001, ****p <
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

5. Fig 2
(a) Western blot analysis was conducted on rat bone marrow mesenchymal stem cells (rBM-MSC) that were initially treated with combinations of H9c2 cardiomyoblast–conditioned media (CM), normoxia, hypoxia, and tumor necrosis factor (TNF)-α (50 ng/mL). Protein expression was normalized to β-actin. Increased expression of (c) angiopoietin-2 (Ang-2), (d) fibroblast growth factor (FGF)-2, (f) vascular endothelial growth factor (VEGF)-1, and (g) transforming growth factor (TGF)-β in TNF-α + H9c2-CM + hypoxia–treated cells was indicated compared with H9c2-CM + hypoxia–treated cells. (b) Ang-1 expression is decreased, whereas (e) FGF-7 expression remains unchanged. Fold differences were normalized to the untreated rBM-MSC group. The error bars on the graphs represent the SD. Statistical significance is at p < 0.05. *p < 0.05, **p < 0.01, ***p < 0.001, ****p <
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

6. Fig 3
Transwell cell migration assay shows that the tumor necrosis factor (TNF)-α + hypoxia–induced secretome enhances the migration of rat bone marrow mesenchymal stem cells (rBM-MSCs) compared with the control secretome (p < 0.05). The TNF-α + hypoxia–induced secretome cell count quantitatively lays closer to the stromal cell–derived factor-1 (SDF-1) positive control than the serum-free Dulbecco’s Modified Eagle’s Medium negative control. The bar graph depicts the cell count per mm2 postmigration in response to each factor. The error bars on the graphs represent the SD.*p < 0.05, ***p < (CM = conditioned medium.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

7. Fig 4
A significant increase in the left ventricular fractional shortening (LVFS) of the rats treated with the tumor necrosis factor (TNF)-α and hypoxia–induced secretome is determined compared with the control secretome (p < 0.01). The error bars on the graphs represent the SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p <
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions

8. Supplemental Fig 1
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2018 The Society of Thoracic Surgeons Terms and Conditions