Figure 2.

Celecoxib / Sorafenib / Sildenafil treatment decreases the expression of the plasma membrane drug efflux pumps ABCB1 and ABCG2. (A–D) OVCAR and SKOV3 cells were treated with vehicle or [sorafenib (2 μM) + sildenafil (2 μM)], and with increasing concentrations of celecoxib (0–4 μM). Six h after drug treatment, cells were fixed in place and immuo-staining performed to detect the expression of ABCB1 and ABCG2 at 10X using a Hermes WiScan wide field microscope. (E and F) Spiky primary ovarian cancer cells and OVCAR established ovarian cancer cells were treated with vehicle or [sorafenib (2 μM) + sildenafil (2 μM) + celecoxib (2 μM)]. Six h after drug treatment, cells were fixed in place and immuo-staining performed to detect the expression of: BAG1, BAG2, BAG3, HSP70, HSP90, GRP78, GRP75, mitochondrial HSP70 and HSP60 using a Hermes WiScan wide field microscope. (G) Spiky primary ovarian cancer cells and OVCAR, CAOV3 and PA-1 established ovarian cancer cells were treated with vehicle or [sorafenib (2 μM) + sildenafil (2 μM) + celecoxib (2 μM)]. Six h after drug treatment, cells were fixed in place and immuo-staining performed to detect the expression of HSP27 using a Hermes WiScan wide field microscope. (H) Spiky and OVCAR cells were transfected with empty vector plasmid (CMV); a plasmid to express GRP78; a plasmid to express HSP27; or to express GRP78 and HSP27. Twenty four h after transfection cells were treated with vehicle or [sorafenib (2 μM) + sildenafil (2 μM) + celecoxib (2 μM)]. Six h after drug treatment, cells were fixed in place and immuo-staining performed to detect the expression of ABCB1 and ABCG2 using a Hermes WiScan wide field microscope. (I) OVCAR and SKOV3 cells were treated with vehicle or [sorafenib (2 μM) + sildenafil (2 μM) + celecoxib (2 μM)] in the presence or absence of elacridar (Pi, 80 nM). Twelve h after drug treatment, cells were processed and subjected to live/dead assays in a Hermes Wiscan system.