Fig. 4

Combination of APG-2449 and paclitaxel inhibits tumor growth of FAK-expressing ovarian cancer xenografts in mice. Western blotting of FAK signaling pathway in PA-1 ovarian cells treated with APG-2449 or defactinib for 4 hours in vitro (A). Efficacy studies in subcutaneous CDX models derived from PA-1 (B; treated for 3 weeks, n = 5 per treatment group) or OVCAR-3 (C; treated for 4 weeks, n = 5 per treatment group; T/C values assessed on Day 28) ovarian cancer cells. D, A mouse trial experiment was conducted in a panel of 6 ovarian cancer PDX models (treated for 3–6 weeks, n = 2 per treatment group). E, Heatmap of genes significantly changed in ovarian PDX tumors that responded to APG-2449 plus paclitaxel combination (synergy ratio > 2) versus nonresponders (synergy ratio < 2). p < 0.05 (false discovery rate < 0.05). F, Western blot analysis of FAK signaling pathways in tumor samples responsive (OV2018) and nonresponsive (OV1396) to APG-2449 plus paclitaxel treatment; tumor samples collected 4 hours after the last administration from the experimental animals shown in panel D. G, IHC staining of CD44- or ALDH1A1-positive cells in OVCAR-3 tumors collected 4 hours after the last treatment from tumor-bearing mice treated with APG-2449, paclitaxel or the combination for 10 days. H, SKOV-3 cells were treated with increasing doses of APG-2449 for 72 hours. I, SKOV-3 cells were treated with APG-2449 (1 μM), paclitaxel (5 nM), or the combination for 72 hours. CD44⁺ and ALDH1A⁺ cells were determined by flow cytometry. ***p < 0.001, **p < 0.01 vs. DMSO controls. Date presented are representative of 2 independent experiments as the mean ± SEM of triplicate biological replicates