Figure 6. Galectin 3 inhibition results in the decrease in RhoA GTPase activity and actin reorganization in HCC cells.
Hepa1-6 cells were transfected with either scrambled or galectin 3 siRNA. RhoA GTPase activity was assessed by pull-down of the GTP- RhoA in non-transfected (NT), scrambled or galectin 3 siRNA-transfected cells. RhoA activation was significantly decreased after galectin 3 siRNA transfection (B, C, mean±SEM, N=4, *p<0.05). The RhoA activation was examined in wt and galectin 3−/− tumors as well (D, E). Less active RhoA was seen in the knockout tumors.
Immunofluorescence staining was performed to assess RhoA (A: b, f, j, green) localization and phalloidin (A: a, e, i, red) to visualize F-actin filaments. RhoA/F-actin was localized to the leading edges in the non-transfected (NT) and scrambled siRNA transfected cells (arrows). The galectin 3 siRNA-transfected cells had a more rounded shape with no lamellipodia formation and the RhoA staining was mainly cytoplasmic (Bar=20 μm).
