Journal: Cell Death & Disease

Article Title: Inhibition of the Hexosamine Biosynthetic Pathway by targeting PGM3 causes breast cancer growth arrest and apoptosis

doi: 10.1038/s41419-018-0405-4

Figure Lengend Snippet: a Schematic representation of HBP. b Schematic drawing of the conversion of FR054 (prodrug) into FR051 (active compound). c Poses of the FR051 (upper), Glc N Ac-6-P (middle), and Glc N Ac-1-P (bottom) molecules in the catalytic cleft of PGM3 and their docking scores (kj/mol). d CETSA curves for PGM3 with FR054 measured in MDA-MB-231 cell extracts from 49 to 70 °C. e ITDRF CETSA curves for FR054 and Glc N Ac-6-P in cell extracts at 58 °C. f HPLC quantification of UDP-Glc N Ac in cell extracts of MDA-MB-231 upon FR054 treatment. All data represent the average ± s.d.; * p < 0.05 (Student’s t -test; comparison with FR054-not-treated sample); N = 3

Article Snippet: Equal amounts of proteins were loaded onto 10% SDS–PAGE gels, transferred to nitrocellulose membranes, and analyzed using the following antibodies: PGM3 (#A304-555A, Bethyl Laboratories, Montgomery, TX, USA; 1:5000), vinculin (#sc-5573, Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA; 1:10000), UAP1 (HPA014659, Sigma-Aldrich; 1:250).

Techniques: Comparison

Journal: Cell Death & Disease

Article Title: Inhibition of the Hexosamine Biosynthetic Pathway by targeting PGM3 causes breast cancer growth arrest and apoptosis

doi: 10.1038/s41419-018-0405-4

Figure Lengend Snippet: Cell death of MDA-MB-231 cells upon treatment with NAGKi and FR054: scheme of treatment ( a ), cell death determined by viable count ( b ), representative images of cells upon treatment ( c ) (4× magnification, 50 μm scale). Efficacy of shRNA silencing of PGM3 enzyme in MDA-MB-231 stable clones, as detected by RNA ( d ) and protein ( e ) levels quantification. f Cell viability of shCTR and shPGM3 clones upon 48 h treatment with FR054 0.5 mM and 1 mM. g , h Cell viability of MDA-MB-231 shCTR and shPGM3 representative clones detected by MTT test ( g ; 48 h FR054 at different doses) and Ann V-FITC staining ( h ; 48 h FR054 0.5 mM). All data represent the average ± s.d.; * p < 0.05, ** p < 0.01 (Student’s t -test; comparison with FR054-not-treated sample); N = 3

Article Snippet: Equal amounts of proteins were loaded onto 10% SDS–PAGE gels, transferred to nitrocellulose membranes, and analyzed using the following antibodies: PGM3 (#A304-555A, Bethyl Laboratories, Montgomery, TX, USA; 1:5000), vinculin (#sc-5573, Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA; 1:10000), UAP1 (HPA014659, Sigma-Aldrich; 1:250).

Techniques: shRNA, Clone Assay, Staining, Comparison