Special AT-rich sequence-binding protein 2 acts as a negative regulator of stemness in colorectal cancer cells

Figure 2 Knockdown of special AT-rich sequence-binding protein 2 promotes adhesion, colony-formation and migration of colorectal cancer cells in vitro. A: SATB2 expression levels in cells infected by virus with different shRNAs targeting SATB2 were detected by Western blot. shRNA named shRNA#1 had the best effect in silencing SATB2 expression in SW480, SW620 and DLD-1 cells; B: Cells in which SATB2 was stably knocked down were isolated into single cells and single cells were cultured for 2 wk to form clones. Then we detected SATB2 expression in different clones by Western blot. We chose SW480/clone7 and DLD-1/clone5 cells to be used in the following assays; C: Adhesion capabilities of cells infected by shRNA#1 virus and its control virus were compared by detecting the cells’ absorbance values to reflect the numbers of adhered cells. Cells with low SATB2 expression had increased adhesion capabilities; D: Abilities of clone formation of CRC cells with stably reduced expression of SATB2 by an infection of shRNA#1 virus were detected to have an increase in colony formation assay; E and F: Transwell chambers were used to detect migration ability of cells with stably reduced expression of SATB2 by an infection of shRNA#1 virus (E) or by culturing the single cells isolated from shRNA#1 virus infected cells to clones (F) and both groups with low SATB2 expression had increased cell migration abilities. Scale bar is 50 μm. Data shown are mean ± SEM. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001 vs control.