Journal: ACS Applied Nano Materials

Article Title: Metal-Coordinated His-Tag Functionalization of Polymeric Nanogels for Therapeutic Applications

doi: 10.1021/acsanm.5c05531

Figure Lengend Snippet: (a, b) NG uptake and apoptosis activation in OVCAR3 cells following the administration of the (a) PAH-HA NGs and (b) PAH-PEG NGs. OVCAR3 cells were incubated for 24 h with medium (CTR) or the indicated Cy5-labeled NGs (0.1 mg/mL) with a 1:1 lys-NTA:His-Rhod molar ratio tag functionalization. At the end of incubation, NG engulfment and caspase-3/7 (Casp3/7) activation were analyzed by flow cytometry. The percentages of Cy5-positive cells and Casp3/7-positive cells were calculated. (c) OVCAR3 cells incubated for 24 h with culture medium (CTR) or the indicated Cy5-labeled NGs. At the end of incubation, the endocytosis of NGs was analyzed by confocal microscopy. Scale bar = 20 μm. Nuclei were stained with DAPI (blue). Relative intensity (RI) of Cy5-positive NGs was quantified in at least 51 cells. Data represent mean ± SEM of triplicate cultures. Statistical significance was determined by Student’s t test. (*) p < 0.05, (**) p < 0.01, (***) p < 0.001, (****) p < 0.0001, ns = not significant.

Article Snippet: The human ovarian carcinoma cell line OVCAR3 (ATCC HTB-161) was obtained from the American Type Culture Collection (LGC Standards, Teddington, UK) and cultured in RPMI 1640 medium (1×), supplemented with GlutaMax Supplement (Cat# 61870036; Thermo Fisher Scientific), 20% fetal bovine serum (Cat# F524-500 ML; Merck KGaA), 10,000 U mL –1 penicillin, and 10 mg/mL streptomycin.

Techniques: Activation Assay, Incubation, Labeling, Flow Cytometry, Confocal Microscopy, Staining