Fig. 3
The nanococktail induced blocked cell growth and elevated apoptosis by triggering DSBs. (A and B) Representative images of A549cisR cell proliferation after treatment with the NC fabricated with Pt-LA2 (5 μM) and LA-SN38 (1 μM) for 24 h. Scale bars, 100 μm. (C and D) Cell cycle distribution of A549cisR cells upon drug treatments for 24 h. Cells were stained with propidium iodide (PI) and subjected to flow cytometry for DNA content analysis. The percentage of the cell population is shown in the histogram. (E and F) Fluorescence images of live/dead cells via calcein-AM/PI staining after A549cisR cells were treated with the optimized NC for 24 h. Calcein-AM stained viable cells, emitting green fluorescence, while PI permeated dead cells and produced red fluorescence. Scale bars, 100 μm. (G and H) Apoptosis analysis of A549cisR cells after exposure to each treatment for 24 h (G). The apoptotic rate was shown in the histogram (H). (I and J) Cells were stained with γH2AX (red) to visualize DNA damage under confocal microscopy. Scale bars, 25 μm. (K and L) Representative images showing the comet assay to quantify the degree of damaged DNA in A549cisR cells. Scale bars, 200 μm. Both tail moment and %tail DNA were analyzed from 50 individual cells with Comet Assay Software Project (CASP) version 1.2.3 (L). The Roman numbers under the axis represent the corresponding treatments: (i) untreated, (ii) Pt(IV)-NP, (iii) SN38-NP, and (iv) NC. The data are presented as the means ± SD; ** p < 0.01, ***p < 0.001