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. 2017 Feb 1;8(11):18031–18049. doi: 10.18632/oncotarget.14949

Figure 3. Knocking-down of Rac proteins reduces the colony formation of U251-tumorspheres.

Figure 3

A. U251-tumorsphere cells stably expressing scrambled shRNA (shScramble) or specific shRNA targeting each Rac (shRac1-3) were counted and 2,500 cells in 2x neurobasal medium were mixed with 0.7% agarose and were seeded in a well of 6-well plate. After culturing for three weeks, sphere colonies were stained with 0.01% crystal violet. Colonies were imaged by Las 3000 Image System and counted by Image J program. B. The quantitated results from at least three independent experiments (p values are derived from Student's t-test. *: p<0.05). C. The examination of the size of spheres. Ten thousand trypsinized U373-tumorsphere cells in 0.35% agarose containing medium were seeded in 12-well and cultured for a week. The tumorspheres formed in the gel were pictured by inverted fluorescence microscope and the area of the sphere was analyzed by Image J program.