Fig. 3

The overexpression of HOXD9 is correlated with malignant biological behavior in GC. a HOXD9 expression was detected by western blot with β-tubulin used as the internal control. b Proliferation in GC cells transfected with vector or HOXD9 was assessed by the colony-forming assay. c Proliferation of GC cells was assessed by CCK-8 assay. **, P < 0.05, ***, P < 0.01, ****, P < 0.001. d DNA synthesis in GC cells was measured by the EdU incorporation assay at 48 h after the indicated transfection. Red fluorescence represents the EdU-positive cells; blue fluorescence is from the Hoechst 33342 stain and represents the total cells. e Scratch wounds were made in monolayer cultures of cells after plating. A quicker closure of the wound in the HOXD9 cells culture from 0 to 72 h based on the wound width is shown in the graph compared to the controls. f Tumor cell invasion capacity was measured with Transwell chambers. Representative image fields of invasive cells on the membrane are shown. g The GC cells were treated with U0126 (20 μM), SP600125 (20 μM) or SB203580 (3 μM) for 48 h. Gene expression was detected by Western blot analysis. β-tubulin was used as the internal control. Scale bars, 100 mm in d