Insulin-like growth factor binding protein-5 influences pancreatic cancer cell growth

Figure 4 Cell cycle distribution of PaC cells that overexpress IGFBP-5 grown in the presence of serum. Stable transfectants (empty vector/Vec-white bar, low/IGFBP-5L-gray bar, high /IGFBP-5H-black bar) were seeded in growth media. After 24 h, the medium was removed and replaced with fresh growth media for 48 h. (A) Cell cycle distribution was determined by propidium iodide staining and FACS analysis. Data shown represent the mean ± SE from 3 experiments performed in duplicate. (B) Cell lysates were prepared for immunoblot analysis of cell cycle regulators and their expression was detected using the appropriate antisera. Membranes were subsequently stripped and reprobed for actin or GAPDH as a load control. Analyses of cyclin E/cdk2 and cyclin B/cdc2 expression were performed simultaneously, therefore, only a single load control, actin or GAPDH, respectively, is shown for each pair. Quantitation of band density represents the level of IGFBP-5-associated expression relative to vector-transfected controls (1.0) normalized to the expression level of actin. ND: Not detected. The results are representative of 3 experiments. ^aP < 0.05, ^bP < 0.01, ^dP < 0.001.