Fig. 3

Effect of GLRX3 inhibition in HPAC and CFPAC-1 pancreatic cancer cells: A Establishment of GLRX3 knockdown cells in HPAC and CFPAC-1 pancreatic cancer cell lines. GLRX3 mRNA and protein levels were downregulated by shRNA transfection. Β-actin and GAPDH served as loading controls; B Cell proliferation was reduced in shGLRX3 transfected cells. The transfected cells (2 × 10³ cells/well) were counted every 24 h using a hemocytometer. The experiment was performed in triplicate and the data are shown as the mean ± SEM; C Colony formation was reduced by GLRX3 knockdown. shControl and shGLRX3 cells were cultured on agar media for 4 weeks. The experiment was performed in triplicate and data are shown as the mean ± SD (p < 0.001). Representative images (0.8x) and graphs were obtained at the end of the experiment; D Formation of spheres was reduced by GLRX3 knockdown. The shControl and shGLRX3 cells (1 × 10³ cells/ml) were cultured in sphere conditioned media on ultralow attachment plates for 7 days. Representative 4x photomicroscope images showed the spheres at 7 days after culture; E shGLRX3 cells formed no or smaller tumors than the shControl cells in vivo. The shGLRX3 and shControl cells were injected into the flank of 6-week old male SCID (HPAC NC and H10 clone) or nude (CFPAC-1 NC and 3B10 clones) mice (n = 5/group) and monitored for 14 weeks. Representative graft images show the results of tumor xenografts at the end of the experiments