Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: Final intracellular copper concentration expressed by ng Cu/mg protein for complexes 1-MPOH and 1-MPSG in c = 1 µM: after 24 h of incubation with cancer or normal lines. Data are expressed as mean ± SEM. The asterisks denote p -values < *0.05, **0.01 compared to control (Two-way ANOVA followed by Bonferroni multiple comparisons test was performed using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Concentration Assay, Incubation, Software

Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: Determination of ( A ) A549 and ( B ) HaCaT cell death modes induced by 1-MPSG: the Annexin V-FITC/propidium iodide (PI) double staining assay (Annexin V-FITC—green fluorescence, PI—red fluorescence) was used to detect phosphatidylserine externalization in apoptosis and analyze the membrane integrity, respectively. FACS quantified apoptosis and necrosis after Annexin V-FITC and PI labelling. The effect of caspase inhibitor (Z-VAD-FMK) on the ( C ) HaCaT cells treated with 1 μM 1-MPSG, ( D ) A549 cells treated with 1 μM 1-MPSG and ( E ) A549 cells treated with 1 μM CDDP. Data are expressed as mean ± SEM. (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Double Staining, Fluorescence, Software

Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: Influence of 1-MPOH and 1-MPSG complexes (c = 1 µM) on the intensity of JC-10 fluorescence in treated A549 cells. Alteration in MMP is given as an emission ratio 570 nm/530 nm. (Control (CTRL)—untreated cells, ciprofloxacin—a negative control, gentamicin—a positive control); * P < 0.05 compared to control). (Two-way ANOVA followed by Bonferroni multiple comparisons test was performed using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Fluorescence, Negative Control, Positive Control, Software

Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: Activation of caspase 3 and caspase 9 in A549 cell line by 1-MPSG . Etoposide (50 μM) and cisplatin (1 μM) were used as positive controls, while untreated cells constituted control (CTRL). Data are expressed as mean ± SEM, * P < 0.5, ** P < 0.1, *** P < 0.01 compared to control. (Two-way ANOVA followed by Bonferroni multiple comparisons test was performed using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Activation Assay, Software

Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: ( A ) IL-6, ( B ) TNF-α production induced by 1-MPSG , ctrl (-) negative “low” control, ctrl (+): positive “high” control and 0.1 μg/ml LPS. Data are expressed as mean ± SEM, * P < 0.05, ** P < 0.01 compared to control CTRL(-). (Two-way ANOVA followed by Bonferroni multiple comparisons test was performed using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Software

Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: ( A ) Photos of A549 cells after 24 h treatment without and with 1-MPSG or pyo (pyocyanin) for cyto-ID hypoxia/oxidative stress detection (λex = 505 nm, λem = 524 nm). ( B ) The ROS production in A549 cells after 30 min., 4, 12 and 24 h using H 2 DCF-DA for: 1-MPOH , MPSG , 1-MPSG , CTRL(+): H 2 O 2 as positive control and CTRL(-): negative control, cells without compound; ( C ) Oxidative stress induced by the total ROS production in A549 cells after 30 min., 4, 12 and 24 h detected by CYTO-ID Hypoxia/Oxidative Stress Test for: 1-MPOH , MPSG , 1-MPSG , CTRL(+): pyocyanin as positive control and CTRL(-): negative control. Data are expressed as mean ± SEM, *P < 0.05, **P < 0.01 compared to control CTRL(-). (Two-way ANOVA followed by Bonferroni multiple comparisons test was performed using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Positive Control, Negative Control, Software

Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images. Data expressed as mean ± SEM); ( E ) Live/dead analysis carried out on spheroids using flow cytometry (spheroids were stained with Propidium iodide and Calcein AM).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Staining, Fluorescence, Concentration Assay, Software, Flow Cytometry

Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: Effect of 1-MPSG on the intracellular ROS generation of ( A ) A549 and ( B ) cells. A549 cells were incubated with various concentrations of 1-MPSG for 24 h. ROS generation was determined using an H 2 DCF-DA probe. Data are presenting as analysis of flow cytometry histograms with the average H 2 DCF-DA fluorescence intensity. Data are expressed as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001 compared to control. (Two-way ANOVA followed by Bonferroni multiple comparisons test was performed using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Incubation, Flow Cytometry, Fluorescence, Software

Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: Changes in fluorescence intensity for A549 cells incubated with 1-MPOH, 1-MPSG at c = 1 µM in presence and absence of NAC (5 mM). The results were obtained after 30 min, 4, 12, and 24 h of incubation using H 2 DCF-DA probe. CTRL(-): as a negative control, cells without copper compounds; CTRL(+): H 2 O 2 . * P < 0.05, compared to control (CTRL(-)). (Two-way ANOVA followed by Bonferroni multiple comparisons test was performed using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Fluorescence, Incubation, Negative Control, Software

Journal: Scientific Reports

Article Title: Evaluation of anticancer activity in vitro of a stable copper(I) complex with phosphine-peptide conjugate

doi: 10.1038/s41598-021-03352-2

Figure Lengend Snippet: Flow cytometry was used to determine the cell cycle distribution of ( A ) A549 and ( B ) HaCaT cells treated with 1-MPSG and cisplatin (CDDP). Data are presented as an analysis of the cell cycle distribution of cells after 1-MPSG and CDDP treatment. Data are expressed as mean ± SEM. (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .).

Article Snippet: Figure 11 ( A ) 3D A549 spheroid model—bright-field confocal image of representative spheroid (left) and the same spheroid stained with Hoechst33342 (blue) and phalloidin-FITC (green) (right); ( B ) 1-MPSG cytotoxicity in A549 and ( C ) HaCaT spheroids—representative live/death fluorescence images of spheroids subjected to 10 μM concentration of 1-MPSG and corresponding control untreated spheroids grown in the same culturing conditions; (The figure was plotted using GraphPad Prism version 5.0.0 for Windows, GraphPad Software, San Diego, California USA, www.graphpad.com .) ( D ) Live/dead spheroids staining—quantification of pixel intensity from confocal images.

Techniques: Flow Cytometry, Software