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. 2017 Aug 15;114(35):E7301–E7310. doi: 10.1073/pnas.1705441114

Fig. 7.

Fig. 7.

Ivermectin (IV) exerts KPNB1-dependent antitumor effects on EOC. (A and B) SKOV3 (Left), CAOV3 (Middle), and OVCAR3 (Right) cells were treated with different doses of IV for 3 d. (A) Cell proliferation/survival was then assessed by WST-1 proliferation (n = 4 each, *P < 0.05 vs. all). (B) Apoptosis was assessed by caspase-3/7 activity in cell supernatants (n = 4 each, *P < 0.05 vs. all). (C and D) SKOV3 cells were treated with vehicle or 16 μM IV for 3 d. (C) Cells were then stained with annexin V and PI, and the population of apoptotic cells was assessed by flow cytometry. (D) Expression levels of cell cycle- and apoptosis-related proteins were assessed by Western blots. (E and F) SKOV3 cells were treated with 8 μM IV. (E) Two days later, cell cycles were assessed by flow cytometry (Left); then, to induce G2/M arrest, cells were treated with 500 ng/mL of NOC for 24 h, and the cell cycles were assayed by flow cytometry (Middle); cells were further incubated for 24 h after withdrawal of NOC and analyzed by flow cytometry (Right). (F) Cell populations at each stage of the cell cycle are shown. (G) SKOV3 cells were transfected with NC siRNA or KPNB1 siRNA. Two days later, they were treated with vehicle or 12 μM IV for 2 d. Cell proliferation/survival was then assessed by WST-1 proliferation (n = 4 each, *P < 0.05). (H–J) SKOV3 cells were treated with vehicle, 2 μM IV, 5 nM PTX, or 2 μM IV + 5 nM PTX for 2 d. (H) Cell proliferation/survival was then assessed by WST-1 proliferation (n = 4 each, *P < 0.05 vs. all) and (I) apoptosis was assessed by caspase-3/7 activity in cell supernatants (n = 3 each, *P < 0.05 vs. all). (J) Cells were stained with annexin V and propidium iodide, and the number of apoptotic cells was assessed by flow cytometry. (K) SKOV3 cells were s.c. injected into immunodeficient NSG mice. One week later, mice were randomly assigned into four groups (control, IV, PTX, and IV+PTX). Mice were then treated with i.p. injection of PBS, or 1 mg/kg of IV and/or 15 mg/kg of PTX 5 d a week for 40 d. Xenograft tumor volumes were then measured at different times after the initiation of treatment (n = 4 each, *P < 0.05 vs. all).