BIRC5 and OCT4 expression in HCC cell lines. (A) Cell lines were cultured in 6-well plates at a density of 105 cells/well for 48 h, and then harvested for measuring expression of BIRC5 and OCT4 by Western blotting. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the loading control, and densitometry analysis was performed to show BIRC5 and OCT4 expression levels normalized with GAPDH density. (B) Cells were harvested, the CD133-positive cells were quantified by flow cytometry and showed in percentages of total cells counted; *, P < 0.05; **, P < 0.01. (C) Cells were infected with Ad5-OCT4 at an MOI of 20 pfu/cell, or transfected with shRNA vectors at 20 μg/well. BIRC5 and OCT4 expression was measured by Western blotting. Densitometry analysis was performed to show BIRC5 expression levels, normalized with GAPDH density; *, P < 0.05; **, P < 0.01. (D, E) The parental, adenovirus-infected and shRNA-transfected HCC cells, including Hep3B (D) and BEL-7404 (E), were cultured in Lab-Tek chamber at a density of 104 cells/well for 48 h, fixed in 4% formaldehyde for 30 min, and labeled by the indicated fluorescent antibodies. DAPI was used to stain cellular nuclei, original magnification 400 × .
Cao et al. BMC Cancer 2013 13:82 doi:10.1186/1471-2407-13-82