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Fig 1.

Structures of the methanolic compounds obtained from the GC-MS analysis.

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Table 1.

Chemical composition of the methanolic extract of O. ficus-indica (L.) Mill. flowers.

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Fig 2.

DPPH assay of the methanolic extract of O. ficus-indica (L.) Mill. flowers.

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Table 2.

MBC and MIC of different strains treated with the methanolic extract of O. ficus indica (L.) Mill. flowers.

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Fig 3.

Cytotoxic activity of (A) O. ficus-indica (L.) Mill. and (B) Paclitaxel as a positive control. The IC50 was evaluated against MCF7, MDA-MB-231, and HEPG2 cell lines.

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Fig 4.

Annexin V/PI assay of the methanolic extract of O. ficus-indica (L.) Mill. flowers against different cancer cell lines.

(A) Dot plots of the flow cytometric analysis of the treated and untreated cells; the quadrant was set to indicate the live cells (lower left corner), early apoptosis (lower right corner), late apoptosis (upper right corner), and necrosis (upper left corner). (B) A graphic bar chart to show the percentages of total apoptosis (early and late apoptosis) of IC50-treated and untreated cells. *** indicated the statistical significance of the results at a P-value <0.001 compared to control untreated cells.

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Fig 5.

Cell cycle assay of the methanolic extract of O. ficus-indica (L.) Mill. flowers against different cancer cell lines.

(A) Histograms of the flowcytometric analysis of the treated and untreated cells; four areas were marked to indicate the cell cycle stages of G0/G1, S, and G2/M phases, in addition to the sub-G1 region. (B) A graphic bar chart to express the percentages of all cell cycle stages of the treated and control cells. (C) A graphic bar chart to express the percentages of cell cycle stages included in the cellular arrest for each cell line.

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Fig 6.

ROS assay of the methanolic extract of O. ficus-indica (L.) Mill. flowers against different cancer cell lines.

DCFDA and flow cytometry were used to assess the oxidative stress in the cells treated with the IC50 concentration of the plant extract. (A) HepG2; (B) MCF-7; (C) MDA-MB-231.

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Fig 7.

Immunoblotting analysis of cells treated with the methanolic extract of O. ficus-indica (L.) Mill. flowers at the IC50 concentration.

(A) Western blot results for each cell line. (B) A graphic bar chart to show the changes in the intensity levels of the studied proteins in the IC50-treated and untreated cells. *** indicated the statistical significance of the results at a P <0.001 compared to control untreated cells.

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Fig 8.

Venn diagram of active metabolites (AM) targets and disease targets [breast cancer (BC) and liver cancer (LC)].

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Fig 9.

PPI network of O. ficus-indica (L.) Mill. used as potential targets for breast and liver cancers.

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Fig 10.

O. ficus-indica (L.) Mill. metabolites target network.

The green polygon is for O. ficus-indica, the blue rectangles are for metabolites, the orange circles are for the top ten ranked targets by the PPI network, and the yellow circles are for the rest.

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Fig 11.

GO enrichment analysis of results for cancer treatment of O. ficus-indica (L.) Mill.

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Fig 12.

KEGG pathway enrichment analysis of results for cancer treatment of O. ficus-indica (L.) Mill.

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Fig 13.

PI3K-Akt pathway.

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Fig 14.

The 2D binding of the active metabolites with a standard PI3K enzyme inhibitor.

A) (A) CID: 20393, (B) CID: 5366701, (C) CID: 520339, (D) CID: 601548, and (E) 2D binding interactions of the co-crystallized ligand of PI3K.

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Table 3.

Docking simulation results of active metabolites against the PI3K enzyme.

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