Fig. 4

Blockage of repopulation of NSCLC cells by MF following CDDP exposure. In a and b, A549 cells and H23, respectively were seeded in six-well plates and allowed to establish exponential growth. On day 3, cells were treated with either CDDP for 1 h or CDDP for 1 h directly followed by chronic treatment with 10 μM MF or 20 μM MF for the indicated times. Cell number was counted at 2-day intervals using microcapillary cytometry. The right side of the panel shows the clonogenic survival assessed by plating 500 viable cells taken from day 15 cultures (A549 cells) or day 36 cultures (H23 cells) in sextuplets and provided normal media for 7 days. At the end of the incubation, cells were fixed with methanol, and stained with crystal violet. Colonies with > 30 cells were counted as positive colonies. Results were analyzed using one-way ANOVA followed by Tukey’s Multiple Comparison Test. *Indicates p < 0.05 compared against vehicle; ^#indicates p < 0.05 compared against CDDP. For A549 cells, CDDP was used at a 100 μM concentration. For H23, CDDP was used at a 40 μM concentration. The lower side of the panels depicts phase contrast images of the treated cultures on day 15 (A549 cells) or 36 (H23 cells)