PMC

(A) Lysates of parental, anti-merlin shRNA (sh-merlin-1 and shmerlin-2), and scrambled shRNA (Scr) SKOV3-IP and OVCAR10 cells were immunoblotted for merlin and GAPDH. (B and C) Anchorage-independent growth of the indicated SKOV3-IP or OVCAR10 cells for 72 h in the presence of DMSO or increasing concentrations of PF-271. Values are means (+/− SEM) of triplicate points. (D and E) Lysates of the indicated cells cultured in suspension with DMSO or increasing concentrations of PF-271 for 72 h were analyzed by immunoblotting for pY397 FAK, total FAK, merlin, and actin.

(A) Anchorage-independent growth of the indicated human ovarian carcinoma cell lines in the presence of DMSO or increasing concentrations of PF-271 for 72 h. Values are means (+/− SEM) of triplicate points (***p < 0.001) and presented as percent of DMSO control. (B) Lysates of the indicated cells cultured in suspension with DMSO or increasing concentrations of PF-271 for 72 h were analyzed by immunoblotting for pY397 FAK, total FAK, and actin. (C) Adherent growth of HEY and OVCAR8 in the presence of DMSO or 0.1 μM PF-271 for 72 h. Values are means (+/− SD) of triplicate points. Lower panels, immunoblotting of adherent cell lysates for pY397 FAK, total FAK, and actin. (D) Lysates of the indicated human ovarian cancer cells cultured in suspension (72 h) were immunoblotted for merlin and GAPDH.

(A) Anchorage-independent growth of 5009 and ID8-IP murine ovarian carcinoma cells for 72 h in the presence of DMSO or PF-271. Values are means (+/− SD) of triplicate points (***p < 0.001). Inset, merlin and GAPDH protein levels in lysates of untreated cells cultured in suspension. (B) Orthotopic tumor growth: representative brightfield and mCherry-fluorescent images of surgically-resected uterine horns (UH), ovarian tumors (T), and kidneys (K) from mice treated with vehicle or PF-271 (30 mg/kg) by oral gavage twice daily (Day 7 to Day 28). Arrows indicate sites of metastasis. (C) Primary 5009 tumor weight in mice treated with vehicle (V, n=11) or PF-271 (n=12). Values are means (± SD). (D) Quantification of peritoneal metastatic tumor sites. Values are means (± SD) (**p < 0.01). (E) Ratio of pY397 FAK phosphorylation to total FAK by densitometry of immunoblotting of lysates from normal ovary, 5009 tumor vehicle-control (V) or 5009 tumor PF-271-treated mice. Normal ovary was set to 1, n = 6 per group (***p < 0.001).