Fig. 1. Identification of Withaferin A as a c-MYB directed therapeutic by Connectivity Map analysis.

a 1309 Connectivity Map (CMAP) drugs ranked based on the significance of regression scores between their transcriptional profiles and that of the query from SPIEDw [33] interrogation of the CMAP database. The Z-score corresponds to the number of standard deviations of the score away from the mean. The position of Withaferin A (WFA, rank 15) is shown, with that of mebendazole [26] (rank 1) for reference. b Western blot example of c-MYB protein expression in THP1 AML cells after 6 h treatment with DMSO or 1 μM WFA. c Volcano plots of RNA-seq analysis showing fold gene expression changes in THP1 cells following treatment with DMSO or 1 μM WFA for 6 h. Expression changes greater than 2-fold and P < 0.05 are shown in red, Wald test. Positions of c-MYB target genes GFI1, MYC, IRF8 and IRX3 are indicated. d GSEA demonstrating enrichment of genes downregulated or upregulated, following peptidomimetic targeting of c-MYB function in AML cells [25], in the WFA induced gene expression changes.