Fig. 6

DCC-2036 inhibits the activity of osteosarcoma cells by activating autophagy (a) The control group was treated with 0 µM, the treatment group received DCC-2036 (2.5 µM), and the combined treatment group was administered DCC-2036 (2.5 µM) alongside 3-MA (3 µM). Osteosarcoma cells were exposed to DCC-2036 (2.5 µM) for 48 h, followed by the addition of 3-MA (30 µM) for an additional 3 h. The growth activity of the three cell groups was assessed using the CCK8 assay at 0, 24, 48, and 72 h. (b-e) Cell treatment and grouping as described in (a). (b) Cells were subjected to three independent experiments to quantify the number of cell colonies. The left graph illustrates a representative result from these experiments, while the right graph presents a statistical analysis of the number of clones. (c) A Transwell assay was utilized to evaluate the invasion and migration of treated 143B cells. The left diagram shows a representative result from three independent experiments, and the right diagram provides a statistical analysis of cell numbers. Magnification was set at 400× with a scale bar of 100 μm. (d) An Annexin V-FITC/PI double dye flow assay was performed to quantify cell death rates. The left graph depicts a representative result from three independent experiments, while the right graph displays a statistical analysis. (e) A Western blot assay was conducted to detect changes in markers related to autophagy