Article Title: CDA directs metabolism of epigenetic nucleosides revealing a therapeutic window in cancer
Figure Lengend Snippet: DNA polymerase and nucleoside kinase activities on modified nucleosides a , MS confirmation of 5hmdC, 5fdC and 5cadC in the purchased nucleosides. b , HPLC-UV chromatogram of nucleosides from DNA extracted from H1299 cells transfected with 5hmdCTP. The abundance of 5hmdC relative to dG is illustrated in the right panel (n=3, standard deviation is shown, n.d.= not detected). c , Coomassie-stained SDS-PAGE gel of recombinant purified DCK and CMPK1 enzymes used in the study. d , Two-dimensional TLC images of DCK reaction products. Dotted lines indicate reference points, which aid in tracking the migration localisation of the nucleosides. The monophosphate in each reaction is circled in red (representative picture, n=3). e , schematic map of nucleoside migration on two dimensional TLC plate (* indicates a background spot coming from ATP and used as a reference point)
Article Snippet: Nucleoside stability Nucleosides were obtained from the following sources: 5hmdC (PY-7588, Berry & Associates), 5fdC (PY-7589, Berry & Associates), 5cadC (PY-7593, Berry & Associates), 5AZAdC (A3656, Sigma Aldrich), ATP solution (Thermo Fisher), [γ-32P] ATP (Perkin Elmer), dC (Sigma Aldrich, D3897), dCMP (Sigma Aldrich, D7625), 5hmdCTP (Bioline, BIO-39046).
Techniques: Modification, Mass Spectrometry, High Performance Liquid Chromatography, Transfection, Standard Deviation, Staining, SDS Page, Recombinant, Purification, Thin Layer Chromatography, Migration