Fig. 5

GINS4 knockdown inhibits lung cancer migration and invasion in vitro. a and b H1299 cells were stably transfected with four distinct GINS4 shRNA expression vectors (shGINS4#1–shGINS4#4) or a control vector (shCTRL). GINS4 expression levels determined by performing (a) western blotting analysis and (b) qRT-PCR are shown. c The MTT assay was performed to assess the viability of H1299 cells stably transfected with two distinct GINS4 shRNA expression vectors (shGINS4#1 and shGINS4#1) or the control vector (shCTRL) (n = 3). d Plate colony formation assay was performed to measure the colony formation ability of GINS4-depleted H1299 cells. Relative colony number is shown as a bar graph (mean ± SD of three separate experiments) in the upper region, and a representative image of the colony number is shown in the lower region (n = 3). e A representative image showing the migration (upper region) and invasion (lower region) of stable GINS4-knockout H1299 cells (n = 3). f Western blotting analysis detected EMT-related proteins in GINS4-depleted H1299 cells (n = 3). g High-content screening and analysis were performed to detect the intensity of E-cadherin and vimentin staining in GINS4-depleted H1299 cells (n = 4). h The GINS4-depleted H1299 cells or control cells were plated in ultra-low-attachment dishes to allow tumor sphere formation (left). The fold changes of spheres formed by both the GINS4-depleted H1299 cells or control cells are shown in the right panel (n = 3). * P < 0.05, ** P < 0.01 and *** P < 0.001