Figure 1

(A) Qualitative assessment of 2YnAd and 6YnAd labelling in MDA-MB-231 cells by in-gel fluorescence scanning. Lanes 1 and 6: molecular weight marker; lane 2: DMSO control; lanes 3, 4 and 5 (1 mM, 0.5 mM and 0.25 mM 2YnAd respectively) and lanes 7, 8 and 9 (1 mM, 0.5 mM and 0.25 mM 6YnAd respectively). (B) Coomassie blue staining of the same gel. (C) In-gel fluorescence scan following the metabolic incorporation, cell lysis, click chemistry and affinity enrichment. Lane 1: DMSO control; Lanes 2, 3 and 4: 1 mM, 0.5 mM and 0.25 mM respectively of 2YnAd was used for the metabolic labelling; lane 5: empty and lanes 6, 7 and 8: 1 mM, 0.5 mM and 0.25 mM respectively of 6YnAd was used for the metabolic labelling. (D) Western blot of the same gel after electrotransfer of proteins onto a nitrocellulose membrane probed using an anti-pan-ADP-ribose antibody (MABE1016, EMD Millipore).