Fig. 2 |. Identification of F₁ subcomplex of ATP synthase as the target of apoptolidin A.

a, Schematic of affinity enrichment workflow and competition experiments used to distinguish specific and non-specific binding partners – (T = [specific] target, N = non-specific binder, U = unbound protein); b, Gel-based profiling of Apop A-PA adducts in MV-4–11 cell or c, K562 cells; d, Volcano plot of identified proteins from TMT multiplexed quantitative proteomics of 1 μM Apop A-PA + vehicle (probe) or 20 μM Apop A (competition) treated MV-4–11 cells in duplicate, adjusted p-values calculated using Limma; e, Illustration of the mitochondrial ATP synthase; f, Immunoblot of MV-4–11 cells treated with Apop A-PA [1 μM] ± Apop A [20 μM] showing specific enrichment of ATP5B, non-specific enrichment of VDAC and TIMM17B, and no enrichment of ATP5G1; g, Inhibition of ATP synthesis after 16 h treatment with Ammo A, Apop A, or Oligm measured in MV-4–11 leukemia cells expressing the ATP:ADP reporter Perceval HR.