Journal: ACS bio & med chem Au
Article Title: Identification of Natural Product Sulfuretin Derivatives as Inhibitors for the Endoplasmic Reticulum Redox Protein ERO1α
doi: 10.1021/acsbiomedchemau.1c00062
Figure Lengend Snippet: A.) PC-9 Cells were subjected to the cellular thermal shift assay in the absence or presence of T151742. Upon addition of T151742, Tagg was quantified for ERO1α, ERO1β, and LSD-1. Quantification of the average of n=3 independent experiments based on western blot analysis for ERO1α, ERO1β, and LSD-1. The dashed line (- - -) line represents the cut-off point at 10% which was used to define our aggregation temperature (Tagg) of individual proteins. B.) MTT viability assay comparing potency between ERO1α inhibitor EN-460, T151742, and T151742 derivative SR-F-114, revealing SR-F-114 is about two-fold less effective in vitro versus T151742 and EN-460 in the PC-9 non-small cell lung cancer cell line. Combined n=3 independent experiments performed in quadruplicates*** p <0.001 by one-way ANOVA. C.) Validation of ERO1α knockout in the PC-9 cell line utilizing CRISPR. D.) Two CRISPR deleted ERO1α clones were subjected to treatment of T151742 as well as the non-targeted control PC-9 cell line. Cellular viability was determined by MTT assay. n=3 independent experiments performed in quadruplicates. **** p<0.0001; by two-way ANOVA.
Article Snippet: EN-460 (Sigma Aldrich, CAT# 328501), LSD-1 Inhibitor IV (Sigma Aldrich, CAT# 489479), and Tranylcypromine (Sigma Aldrich, CAT # 616431) were purchased.
Techniques: Thermal Shift Assay, Western Blot, MTT Viability Assay, In Vitro, Knock-Out, CRISPR, Clone Assay, Control, MTT Assay
