Journal: NAR Cancer
Article Title: EHF is a novel regulator of cellular redox metabolism and predicts patient prognosis in HNSCC
doi: 10.1093/narcan/zcac017
Figure Lengend Snippet: EHF Represses Genes Involved in Drug and Glutathione Metabolism. ( A ) Expression of genes associated with xenobiotic metabolism from Figure , in EHF overexpression (A253 and SCC25) and EHF depleted (CAL-27) cells. Genes missing from any heatmap are not expressed in that particular cell line. ( B ) Western blot validation of the repression of AKR1C1 and AKR1C2 by EHF. ( C ) Expression of genes associated with glutathione metabolism from Figure and , in EHF overexpressing (A253 and SCC25) and EHF depleted (CAL-27) cells. ( D ) Western blot showing the expression of SLC3A2 and GPX2 following the overexpression and depletion of EHF. ( E ) Quantification of reduced and oxidized glutathione ratios in all three cell lines, showing that EHF expression results in the loss of reduced glutathione (GSH). Experiment was performed in triplicates, and measurements were taken twice. The bar graphs are presented as average values ± standard deviation (error bars).
Article Snippet: Immunoblotting was performed using the following antibodies: EHF (5A5.5); SOX2 (D6D9, Cell signaling technologies); GAPDH (MAB374, EMD Millipore); β-Tubulin (MAB3408, EMD Millipore); AKR1C1 (CPTC-AKR1C1-1, DHSB); AKR1C2 (CPTC-AKR1C2-1, DHSB); NRF2 (D1Z9C, cell signaling technologies); ELF3 (MAB5787, R&D), SLC3A2 (15193-1-AP, Proteintech) and GPX2 (MAB5470-SP, R&D).
Techniques: Expressing, Over Expression, Western Blot, Biomarker Discovery, Standard Deviation