Figure 3.

High-Throughput Screen Identification of the miR-34a Synergistic Small Molecule Ouabain

(A) Data show XY plots with cell counts and activated caspase 3 + cells in doxycycline-treated (DOX+) versus control cells (DOX−). Data summarized from 9,850 small molecules of biologically known activity.

(B) Plot of fold change in total cell counts and caspase 3 + cell counts from cells treated with doxycycline compared with control untreated cells. Data from the top 50 molecules with decreased cell numbers and increased caspase 3 + cells are shown.

(C) Synergy plot of miR-34a and ouabain-treated A549 and HCC827 lung cancer cells. Lowe's additivity combination index was calculated and plotted as heatmap using Combenefit data analysis tool.

(D) MTT assay-based validation of the observed synergy with ouabain and miR-34a in A549, HCC827, H1975, and CALU6 lung cancer cells. Data show % cytotoxicity in 40 nM miR-34a treated (blue line) versus 40 nM miR-control (black line) with an increasing dose of ouabain. Data were fitted by non-linier regression with variable slope. Goodness of fit showed R squared for all the IC50 plots greater than 0.9.