JK184 inhibits growth of claudin-low cell lines. (A) Real-time RT-PCR analysis of Gli1 RNA expression in JK184-treated claudin-low cell lines. Doses were 0, 0.002, and 0.004 μM JK184 for 72 h. Values were normalized to DMSO vehicle control, error bars are the standard error between two independent experiments. * = P ≤0.05, t test. ** = P ≤0.005, t test. (B) Western blot showing protein expression levels in claudin-low cells treated as in A). Lanes 19 to 21 are from a separate gel. (C) Dose-response curve for 72 h JK184 treatment of claudin-low and MCF10a, HMLE-shGFP, HMLE-pBP, and MTSV1-7 cell lines. (D) Flow cytometric analyses of cells treated with JK184 (0.02 μM) for four days followed by staining with Annexin-V and propidium iodide (PI). Images representative of three independent experiments. (E) Light microscopy images of cells treated as in A). Scale bar indicates 100 μm.