Figure 3
From: A cellular screen identifies ponatinib and pazopanib as inhibitors of necroptosis
Chemical proteomics identifies necroptosis pathway members as targets of ponatinib. (a) Structure of ponatinib and the analog c-ponatinib used for affinity purification. (b) Cell viability was determined in FADD-deficient Jurkat cells treated overnight with 10 ng/ml TNF-α, 10 μM Nec-1 and ponatinib (P) or c-ponatinib (c-P) as indicated. Cell viability was assessed using a luminescence-based readout for ATP (CellTiter Glo). Data represent mean value±S.D. of two independent experiments performed in triplicates and normalized to untreated control. (c) Proteins identified in mass-spectrometry-based affinity purification experiment with c-ponatinib. The x-axis of the bubble plot represents the statistical significance (P-value) of protein enrichment over the competition assay with free ponatinib estimated using the modified Decontaminator method (see Materials and Methods), and the y-axis the reduction in spectral counts (%) upon competition. Bubble size is proportional to the average spectral counts in non-competed condition. Dashed line indicates the P-value threshold (0.05)
