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. 2019 Apr 1;8(4):301. doi: 10.3390/cells8040301

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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The effect of d-NGF(1-15) and NGF(1-14) peptides on copper homeostasis. In (A): representative merged confocal microscopy images of coppersensor-1 (red) and Hoechst33342 ( blue) stained cells after 15 min of treatment with 10 μM NGF(1-14) or d-NGF(1-15), either in the absence or in the presence of 1 µM CuSO₄. Untreated and 1 µM CuSO₄ treated-cells are included as negative and positive controls, respectively (scale bar = 10 μm). (B,C): Quantitative analysis of the confocal micrographs for total cytoplasmic Cu⁺ (B) and nuclear Cu⁺ (C) in the cells incubated with the peptides in basal medium (, ) or copper-supplemented medium (, ). In (D,E): quantitative analyses of total intracellular copper (D) or nuclear-confined copper (E) after PC12 cells incubation for 10, 15, 20 or 30 min with 10 μM NGF(1-14) (cytoplasm: ; nuclei: ) or 10 μM d-NGF(1-14) (cytoplasm: ; nuclei: ). ** p < 0.01, *** p < 0.001, versus control.; ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus the preceding incubation time; § p < 0.05, §§§ p < 0.001 versus [NGF(1-14)].

Inline graphic — The effect of d-NGF(1-15) and NGF(1-14) peptides on copper homeostasis. In (A): representative merged confocal microscopy images of coppersensor-1 (red) and Hoechst33342 ( blue) stained cells after 15 min of treatment with 10 μM NGF(1-14) or d-NGF(1-15), either in the absence or in the presence of 1 µM CuSO₄. Untreated and 1 µM CuSO₄ treated-cells are included as negative and positive controls, respectively (scale bar = 10 μm). (B,C): Quantitative analysis of the confocal micrographs for total cytoplasmic Cu⁺ (B) and nuclear Cu⁺ (C) in the cells incubated with the peptides in basal medium (, ) or copper-supplemented medium (, ). In (D,E): quantitative analyses of total intracellular copper (D) or nuclear-confined copper (E) after PC12 cells incubation for 10, 15, 20 or 30 min with 10 μM NGF(1-14) (cytoplasm: ; nuclei: ) or 10 μM d-NGF(1-14) (cytoplasm: ; nuclei: ). ** p < 0.01, *** p < 0.001, versus control.; ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus the preceding incubation time; § p < 0.05, §§§ p < 0.001 versus [NGF(1-14)].