FIG. 3.

Effect of K8.1 and/or gB inhibition on VEGF production. (A) BCBL-1 cells were transfected with plasmid vectors expressing nonspecific siRNA, ps5 (K8.1 target), or ps5/K8.1co. Cells were induced with TPA (+) or left uninduced ([) starting at 24 h posttransfection. Cell pellets were collected 16 h postinduction, and total RNA was extracted. First-strand cDNAs were produced and amplified by PCR using primers described in Table 1, and the resultant 250-bp DNA fragments were quantified by densitometry and normalized to GAPDH. (B) Experiments similar to those described for panel A using plasmid vectors expressing nonspecific siRNA, ps18, ps18+gBco, and ps18+gBR. (C) Experiments similar to those described for panels A and B with plasmid vectors expressing nonspecific siRNA, ps5/s10, ps18+ps22, and ps5/s10 + ps18-22. Cells were induced with TPA (+) 24 h posttransfection or left uninduced (−), supernatants were collected 48 h postinduction, and the total amount of protein was determined. Equal amounts of protein from the supernatants were loaded on SDS-PAGE gels. Experiments were conducted in triplicate, and error bars represent SEM. The bar graphs represent the densitometric analysis results obtained using the Image J software.