Figure 5. IMD-0354 Treatment Inhibits Melanoma Cell Growth, Induces Cell Cycle Arrest, Apoptosis and Autophagy.

(a) A431 and A375 cells were treated with the indicated concentrations of IMD-0354. Cell viability was measured by ATPlite after 72h. (b) A431 and A375 cells were plated at low density (500 cells/well in 6-well plates) and grown in medium containing indicated concentrations of IMD-0354. The number of colonies formed after 10 days in culture was determined by crystal violet staining (left) and quantified (right). (c) A375 cell spheroids were treated with indicated concentrations of IMD-0354. Images were taken under bright field microscope after three days (left) and quantified (right). (d) A375 cells were treated with indicated concentrations of IMD-0354 for 24h. Cells were stained with propidium iodide (PI) and cell cycle was analyzed by FACS.

(e) A375 cells were treated with IMD-0354 (2 μM) for 24h. Cells were stained with anti-LC3 and DAPI. Scale bar, 20 μm. (f) A375 cells were seeded in transwell and treated with IMD-0354 (0.25 μM or 1 μM) for 16h. Migrated cells were visualized by crystal violet staining. Statistical analysis was performed by one-way ANOVA for the comparison of more than two groups. Unpaired t-test was used for the comparison of two groups. Data are shown as the mean ± SD, n = 3. **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001.