Fig. 1: Chemical compound screen discovers that plicamycin and everolimus specifically target MDST8.
A Scheme of the screening campaigns. B MDST8 or LoVo cells were treated with 71 drugs in the anticancer library at a concentration of 0.1 μM for 72 h. The percentage of AnnexinVhighDAPIhigh cells was measured by flow cytometry as an indicator for cell death. Each parameter depicts the mean value of three times repeated experiments and is depicted in a hierarchically clustered heatmap. The blue and red tiles in the heatmap represent the percentage of Annexin VhighDAPIhigh death cells range from 0 to 100%. C Identification of plicamycin, everolimus, and rapamycin as chemicals that specifically kill MDST8 but not LoVo cells. MDST8-RFP or LoVo-GFP cells were treated with 71 drugs in the anticancer library at the concentration of 0.1 μM for 72 h. Debris and cells depicting nuclear pyknosis were excluded, and healthy cells were enumerated. The untreated control was normalized to 1. D Results reported in a bi-parametric plot, showing the normalized healthy cell counts after treatment comparing between MDST8-RFP and LoVo-GFP. E Images show representative pictures of colonies formed as observed upon crystal violet staining after treatment of MDST8 and LoVo cells with 10 nM everolimus (EVE), 10 nM plicamycin (PLI), or 2 μM sunitinib (SUN) for 3 to 4 weeks. F The bar chart represents the number of clones with a size greater than or equal to 50 μm2. Error bars indicate SEM. Asterisks refer to significant effects for treatments versus control (Ctrl) (paired Student’s t test, ***P < 0.001).
