JK184 is more effective at inhibiting EMT cell proliferation. (A) Clustering of Spearman ranked data of inhibition concentration (IC) values for the inhibitor screen. Rank values are as follows for μM concentrations: 1: no IC values are between 0 and 10; 2: IC10 between 0 and 10, and IC25 not between 0 and 10, and IC50 not between 0 and 10; 3: IC25 between 0 and 10, and IC50 not between 0 and 10; 4: IC50 between 0.1 and 10; 5: IC50 between 0.01 and 0.1; 6: IC50 between 0.001 and 0.01; 7: IC50 < 0.001. Asterisks mark the agents mentioned in the text. (B) Dose-response curve for JK184 treatment of HMLE-shEcad cells (blue) and HMLE-shGFP cells (green). Error bars indicate coefficient of variance between triplicate treatment. (C) Dose-response curve for JK184 treatment of HMLE-shEcad cells (blue), HMLE-shGFP cells (green), HMLE-Snail cells (purple), and HMLE-pBP cells (gray). Treatment was conducted in 96-well plates. (D) Immunoblot showing response of GLI1 expression in HMLE cells with JK184 drug treatment. Doses were 0, 0.002, and 0.004 μM JK184 for 72 h. GAPDH serves as a loading control. GLI1 quantification levels were normalized to HMLE-shEcad untreated levels, and appear below the lane numbers in the figure. (E) Real-time PCR data showing relative GLI1 mRNA transcript levels from cells treated as in D). Error bars represent standard error between two independent experiments. * = P ≤0.05, t test. ** = P ≤0.005, t test. GAPDH, glyceraldehyde phosphate dehydrogenase; GLI1, glioma-associated oncogene 1; HMLE, human mammary epithelial cells, transformed with large T and small t antigens.