Journal: Scientific Reports
Article Title: Selective uptake of epidermal growth factor-conjugated gold nanoparticle (EGF-GNP) facilitates non-thermal plasma (NTP)-mediated cell death
Figure Lengend Snippet: Preparation of the Epidermal Growth Factor (EGF) conjugated gold nanoparticle (GNP). ( A ) The experimental scheme shows conjugation between EGF and GNP. Three different linkers were used for conjugation; MPA (3-mercaptopropionic acid), MUA (11-mercaptoundecanoic acid), and MHDA (16-mercaptohexadecanoic acid) ( B ) Serum-starved (24 hours) A549 cells were treated with 1% BSA, EGF (100 μg/ml), 5 nm GNP, 5 nm GNP-MPA-EGF, 5 nm GNP-MUA-EGF, and 5 nm GNP-MHDA-EGF respectively. Whole-cell extracts were analyzed by Western blotting for assessment of the level of phospho-ERK protein. 14-3-3ξ was used as a loading control. Quantification of Western blots was performed with Image J. All density values were normalized with the loading control. ( C ) Whole-cell extracts were analyzed by dot blots. EGF antibody detected 5 nm GNP-MUA-EGF and 5 nm GNP-MHDA-EGF. The experiment was performed independently at least three times. ( D ) Serum-starved (24 hours) A549 cells were treated with 1% BSA, EGF (100 μg/ml), 5 nm GNP, 5 nm GNP-MUA-EGF, and 5 nm GNP-MHDA-EGF for 6 hours at 37 °C, respectively. MTT assay was performed to assess cell viability after incubation without plasma irradiation. ( E ) Serum-starved (24 hours) A549 cells were treated with EGF (100 μg/ml), 5 nm GNP, 5 nm GNP-MUA-EGF, and 5 nm GNP-MHDA-EGF, respectively. Cells were incubated at 37 °C or 29.5 °C for 6 hours. EGFR was stained with green fluorescent antibody, and DNA was stained with Hoechst 33342. The scale bar on each image represents 10 μm.
Article Snippet: Briefly, 0.1 mg/ml aqueous solution of 3-mercaptopropionic acid (MPA), 11-mercaptoundecanoic acid (MUA), and 16-mercaptohexadecanoic acid (MHDA) were prepared and incubated with 5 nm colloidal gold (Sigma-Aldrich, St. Louis, MO) suspension.
Techniques: Conjugation Assay, Western Blot, MTT Assay, Incubation, Irradiation, Staining