Journal: Oncotarget

Article Title: Vorinostat suppresses hypoxia signaling by modulating nuclear translocation of hypoxia inducible factor 1 alpha.

doi: 10.18632/oncotarget.18125

Figure Lengend Snippet: Figure 1: SAHA suppresses HIF-1α and HIF-2α induction in response to hypoxia. A. Hep3B cells were exposed to hypoxic conditions (1% O2) for 16 hr in the presence of 0.5 or 1 μM of SAHA (+, and ++, respectively). Representative western blot with normalized densitometric values (protein/actin loading control) show decreases in HIF-1α and HIF-2α expression upon exposure to SAHA. B. Luciferase reporter assay demonstrates significant decreases (p < 0.0001) in HRE-associated luciferase activity in response to SAHA under hypoxic conditions. C. Effects of SAHA on hypoxia related gene expression in Hep3B cells exposed to SAHA for 16 hr under conditions of 21% or 1% O2 analyzed by qRT-PCR, showing significant suppression of EDN1, EPO, GLUT1, and VEGFA in response to both 0.5 and 1 μM SAHA with no significant change in HIF1A or HIF2A expression. D. Tumor cell lines U87 MG, U2OS, and MG63 were exposed to 0.5 μM SAHA for 16 hr under 21% or 1% O2, with resulting HIF-1α and HIF-2α suppression similar to that observed in Hep3B cells. Representative western blot with normalized densitometric values (protein/actin loading control) are shown. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The primer sets used in the present study were: EDN1 (Origene HP205717), EPO (Origene HP200740), GLUT1 (Origene HP209446), VEGFA (QIAGEN QT01682072), and ACTB (Promega G5740).

Techniques: Western Blot, Control, Expressing, Luciferase, Reporter Assay, Activity Assay, Gene Expression, Quantitative RT-PCR

Journal: Oncotarget

Article Title: Vorinostat suppresses hypoxia signaling by modulating nuclear translocation of hypoxia inducible factor 1 alpha.

doi: 10.18632/oncotarget.18125

Figure Lengend Snippet: Figure 6: SAHA suppressed hypoxia signaling in Hep3B cell xenografts in vivo. A. In-vitro luciferase assay shows decreased HRE activity in Hep3B HRE-Luc cells in the presence of SAHA. B. Tumor growth curves of Hep3B HRE-Luc cell xenografts treated with SAHA show significantly decreased tumor size at 7- and 9-days post-implantation (p < 0.05, p < 0.01, respectively). C. Average tumor weight of excised xenografts is significantly reduced in SAHA-treated mice compared to control mice (p < 0.05) at study endpoint. D. Example luminescence of excised Hep3B HRE-Luc cell xenografts at study endpoint. E. HRE-luc luminescence is significantly decreased in SAHA treated mice compared to control (p < 0.05) at study endpoint. F., G. VEGFA and EPO expression is significantly decreased in tumors resected from SAHA treated mice at study endpoint. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The primer sets used in the present study were: EDN1 (Origene HP205717), EPO (Origene HP200740), GLUT1 (Origene HP209446), VEGFA (QIAGEN QT01682072), and ACTB (Promega G5740).

Techniques: In Vivo, In Vitro, Luciferase, Activity Assay, Control, Expressing

Journal: Journal of Translational Medicine

Article Title: Manufacturing mesenchymal stromal cells in a microcarrier-microbioreactor platform can enhance cell yield and quality attributes: case study for acute respiratory distress syndrome

doi: 10.1186/s12967-024-05373-7

Figure Lengend Snippet: Protein Expression in gelatin microcarrier-microbioreactor and T25 Flask. The term ‘therapeutic’ in green font signals an anticipated correlation with improved therapeutic efficacy in ARDS, as hypothesized based on review of existing literature. The term ‘Pro-inflammatory’ in red font signals an anticipated correlation with an anti-therapeutic effect in ARDS, as hypothesized by review of existing literature. MSC gene expression was evaluated at the protein level for a subset of genes from Fig. to confirm protein translation and evaluate the absolute concentration of effector molecule present. Protein-level data corroborated some – though not all – of the trends reported from mRNA-level data. Protein concentration was evaluated from MSC secretome by harvesting the conditioned media from culture days 5–7 and utilizing colorimetric ELISA for each analyte. Analytes were assessed at two dilutions, each in triplicate wells, for each condition. Protein concentration was normalized to cell count. Conditioned media was vortexed with 0.02% Tween20 to rupture liposomes prior to starting the ELISA protocol. Remaining steps followed the manufacturers’ protocols. Significance bars indicate results of unpaired two-tailed t-tests. *, p < 0.05. GMC-MB: Gelatin microcarrier-microbioreactor

Article Snippet: Colorimetric sandwich ELISA kits (R&D Systems, DY286-05, DY6030B-05; Thermo Scientific, 88-7176-86, BMS2034MST; Fisher Scientific, CHC1323) were used to quantify secretome protein expression.

Techniques: Expressing, Concentration Assay, Protein Concentration, Enzyme-linked Immunosorbent Assay, Cell Counting, Liposomes, Two Tailed Test