Figure 4. Induction of apoptosis and autophagy in response to EGFR-targeted C225-NP in NSCLC cells.

(a) Percentage of NSCLC cells treated with different doses (0.6, 1.2 and 2.4×10¹⁰ particles) of IgG-NP or C225-NP for 72 hrs that was in the sub-G₁ phase of the cell cycle. This percentage was determined by DNA flow-cytometric analysis. Untreated cells and cells treated with C225 antibody alone served as control. A dose-dependent increase in the number of HCC827 cells in subG1 phase was observed in both IgG-NP and C225-NP treatments. However, the increase in number of cells in subG1 was significantly higher in C225-NP treatment than in IgG-NP (^*P<0.05). There was no marked increase in subG1 phase in H520 cells between IgG-NP- and C225-NP-treatments. (b) Detection of GFP-LC3 dots indicative of autophagy on NSCLC cells that were not treated or treated with C225 antibody, IgG-NP or C225-NP (3×10⁹ particles) for 72 hrs on chamber slides. Scale bar = 50 µm (c) Quantification of the number of cells with GFP-LC3 dots on untreated and treated NSCLC cells. The number of cells with GFP-LC3 dots was higher in C225-NP-treated HCC827 cells compared to all other treatment groups. In H520 cells there was no increase in the number of GFP-LC3 dots when treated with C225-NP and compared to all other treatment groups. Results shown are the means ± S.D. of three independent experiments. *P-value <0.05 vs untreated control, C225 antibody, and IgG-NP on HCC827 cells.