Fig. 3

a–f CsA enhances the cytotoxicity of Gef when co-delivered by NPs to three NSCLC cell lines by promoting apoptosis. Cells were treated with Gef in the absence or presence of CsA via NPs for 48 h before being stained with Annexin V (AV) and propidium iodide (PI), and the apoptotic rates were determined by flow cytometry. The quantitative apoptotic rates are displayed in the upper panel, and the proportions of apoptotic cells are shown in the lower panel. g–i LIVE/DEAD staining was analyzed by confocal microscopy. PC-9, PC-9-GR, and H1975 NSCLC cells were treated with CsA-NPs (1, 1, 1 μM), Gef-NPs (1, 20, 10 μM), and CsA/Gef-NPs (1 + 1, 1 + 20, 1 + 10 μM). Untreated cells were used as the control. Live cells were stained with calcein-AM, while dead cells were stained with PI. Fluorescence images of the same samples were captured at 490 nm (green) for the Calcein-AM signal and at 545 nm (red) for the PI signal and merged into new images. Scale bars represent 100 μm. j–l The death rates of PC-9, PC-9-GR and H1975 cells were analyzed. The difference in the death rates of cells treated with CsA-NPs or Gef-NPs alone or in combination was significant