Extraction scheme for plant powder.

HeLa cells were treated with the indicated extracts at various concentrations (1, 5, 10, 50, 100, 250, 500 and 1000 µg/mL) and MTT survival tests were performed. The toxicity curves of the ethanol extracts of Mahonia bealei, Ficus semicordata, Gnetum montanum, Crinum asiaticum, Mallotus barbatus, Aganope balansae, Hedyotis capitellata, Anisomeles indica and Paclitaxel are presented. The cell viability is shown as % relative to untreated control. Cell viability curves were drawn using GraphPad Prism software.

The MCF-7 cells were treated with the extracts at various concentrations of 1, 5, 10, 50, 100, 250, 500 and 1000 µg/mL. The toxicity curves of the ethanol extracts of Mahonia bealei, Ficus semicordata, Gnetum montanum, Crinum asiaticum, Mallotus barbatus, Aganope balansae, Hedyotis capitellata, Anisomeles indica and Paclitaxel were presented. The cell viability is shown as % relative to untreated control. The curves were drawn by using GraphPad prism software.

Effect of AI-EtE on development and survival of zebrafish embryos at different time points. The effects of AI-EtE at various doses (0, 5, 10, 12.5, 25, 50, 75, 100, 200 and 400 mg/L) on defects during development (A), lethality (B), and hatching percentage (C) of zebrafish embryos at 24, 48, 72 and 92 hpf, respectively, are presented as dose-response curves. Representative images of 96 hpf zebrafish larvae presenting various developmental defects upon AI-EtE exposure at different doses, (D) 0 mg/L (negative control), (E) 25 mg/L, (F) 50 mg/L, (G) 75 mg/L, (H) 100 mg/L, (I) 200 mg/L, and (J) 400 mg/L. Illustrated defects on larvae include yolk-sac oedema (e), haemovascular defect (h), and necrosis (n) as indicated with arrows.

HeLa cells were treated with DMSO (negative control), AI-EtE at IC50 dose of 38.8 mg/L or paclitaxel (positive control) at IC50 dose of 13.5 ng/mL for 24 h before collecting, fixing, staining with propidium iodide (PI)/annexin V-FITC and subjecting to flow cytometry system to measure of apoptosis. Representative data sets of apoptotic pattern of HeLa cells upon DMSO treatment (A), AI-EtE treatment (B), and paclitaxel treatment (C) are presented. Transcript levels of BAX (D), CASPASE-8 (E), CASPASE-3 (F) in HeLa cells treated with AI-EtE and paclitaxel for 12 h and 24 h. *, ** and ***, significant differences with p values < 0.05, <0.01 and <0.001, respectively. TBP was used as internal control gene. NC: negative control, PC: positive control (paclitaxel).

HeLa cells were treated with DMSO (negative control), AI-EtE at IC50 dose of 38.8 mg/L or paclitaxel (positive control) at IC50 dose of 13.5 ng/mL for 24 h before collecting, fixing, staining with propidium iodide and subjecting to flow cytometry system to measure of cellular DNA content. Representative data sets of cell cycle pattern of HeLa cells upon DMSO treatment (A), AI-EtE treatment (B), and paclitaxel treatment (C) were presented. Transcript levels of CDKN1A (D), CDKN2A (E) and p53 (F) in HeLa cells treated with AI-EtE and paclitaxel for 12 h and 24 h. *, ** and ***, significant differences with p values <0.05, <0.01 and <0.001, respectively. TBP was used as internal control gene. NC: negative control, PC: positive control (paclitaxel).

Anchorage-independent growth of HeLa cells. Colonies formed on the soft agar in case of control cells (A,C) and AI-EtE-treated cells (B,D) at magnifications of 2X (A,B) and 5X (C,D). The differences in colony formation ability on soft agar of HeLa cells are presented in a graph (E). *, significant differences with p values < 0.05.