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. Author manuscript; available in PMC: 2016 Jun 1.
Published in final edited form as: Biochim Biophys Acta. 2015 Mar 6;1848(6):1308–1318. doi: 10.1016/j.bbamem.2015.03.001

Figure 4.

Figure 4

CL-DNA NPs colocalize with Rab5-GFP-labeled endosomes. (A, B) Fluorescent micrographs and cell outlines of wildtype Rab5-GFP expressing L-cells that have been incubated with fluorescent RGDtagged NPs. The three overlayed channels represent lipid (red), DNA (blue) and Rab5-GFP (green). In (A) Low-σM refers to NPs with DOTAP/PC/RGD-PEG2K-lipid at a molar ratio of 30/60/10. The cells in (B) were incubated with High-σM NPs composed of MVL5/PC/RGD-PEG2K-lipid at a molar ratio of 50/45/5. (C, D) High magnification of boxed regions from (A, B) show intracellular complexes with (ii, iii, v) and without (i, vi) Rab5 colocalization. (E, F) Intensity profiles (dashed lines in C, D) showing signals from all three fluorescent channels. Early endosomes lacking NPs are also observed (iv). (G, H) Quantitative colocalization shows that a statistically significant difference in the number of NPs found with (+) and without (−) Rab5-GFP at 30 and 60 minutes respectively. Although an increase in the number of NPs lacking colocalization with Rab5-GFP is observed from 30 to 60 minutes. The number of intracellular NPs colocalized with Rab5-GFP does not significantly change. All scale bars are 10 µm. Student t-test for colocalization * P < 0.025, ** P < 0.001.