Fig. 3

Cytotoxicity assessment of AM in HeLa cells. a The dose-dependent cytotoxicity of AM was assayed at concentrations ranging from 0.39 to 100 µg/mL against HeLa and OUMS-36 cells and expressed as percent inhibition of cell viability of treatment groups with AM compared with that of the non-treatment control group. X axis, the logarithm of the AM concentration (µg/mL) to the base 10. n = 3. Error bar, mean ± SD. b The time-dependent cytotoxicity on HeLa cells of AM at IC₅₀ (square, 2.6 µg/mL) and IC₉₀ (triangle, 6.25 µg/mL) concentrations for various times (0, 4, 8, 12, 24, 36, 48, and 60 h). Non-treatment group was used as a control (circle). Data are expressed as relative cell viability at each time to that at time 0. c Representative images of the clonogenic assay. HeLa cells were treated with AM at IC₅₀ (2.6 µg/mL) and IC₉₀ (6.25 µg/mL) concentrations for 36 h and then cultured in a compound-free medium for 10 d. Subsequently, the cells were stained with crystal violet and observed under a microscope. d Numbers of colonies containing over 50 cells formed by the 10-d culture in the clonogenic forming assay in panel (c). Data are represented as mean ± SD and statistically analyzed by one-way analysis of variance (ANOVA); Statistically significant differences were observed between the treated groups and the non-treated group; ‡, p < 0.0001; n = 3