MSP58 depletion suppressed both anchorage-dependent and independent colony formation. (A, B) Equal numbers of U251-S cells, Parental U251 cells and vector control U251 cells (U251-H1 neo and U251-NC) were seeded onto 60mm dish. After 14 days, the cells were fixed and stained with Giemsa (A). The average number of colonies formed in three independent experiments was quantitated (B). (C, D) Plate colony formation ability of clone U251-Smix was detected (C) and the average number of colonies formed in three independent experiment were calculated (D). (E, F) Equal numbers of U251-S cells, parental U251 cells and vector control U251 cells (U251-H1 neo and U251-NC) were plated in 0.3% soft agar and cultured for 14 days. Colony formation was photographed under microscope (E) and scored (F). Clone numbers were mean ± SD of three independent experiments. *P < 0.05 versus U251, U251-H1 neo and U251-NC control cells.