PAP-3 increased ROS production in MCF-7 cells.

<p>(A) Cells were treated with or without 200 µg/mL PAP-3 for 0, 6, 12, and 24 h, and the ROS levels were measured by both the Laser Scanning Confocal Microscopy system and flow cytometry after an incubation with DCF-DA (for H₂O₂) or DHE (for O₂^•−) fluorescence probe stain. (B) SOD and NAC eliminated PAP-3-induced ROS generation. Cells were protected by SOD (30 U/mL), and NAC (1 mM) with 1 h pretreatment when co-incubated with 200 µg/mL PAP-3 for another 24 h, and the ROS levels were stained with DCF-DA or DHE. (C) Impact of SOD (30 U/mL) and NAC (1 mM) on the viable cell was determined by trypan blue dye exclusion assay. Cells were pretreatment with SOD (30 U/mL), and NAC (1 mM) for 1 h, then co-treated with 200 µg/mL PAP-3 for another 48 h. (D) Impact of SOD and NAC on the apoptotic value was determined by Annexin V-FITC/PI staining. All experiments were done independently in triplicate per experimental point, and representative results are shown.</p

PAP-3 induced mitochondrial apoptotic pathway.

<p>(A) The protein expression was determined with or without 200 µg/mL PAP-3 by Western blot, with GAPDH as loading control. (B) Evaluation of mitochondria membrane potential in MCF-7 cells was done with Rhodamine 123 stain by flow cytometry. The percentage of M1 reflects the reduction of ΔΨm. All experiments were done independently in triplicate per experimental point, and representative results are shown.</p

The HPLC chromatograms of standard monosaccharides (A) and component monosaccharides released from PAP-3 (B).

<p>PAP-3 was hydrolyzed into component monosaccharides with trifluoroacetic acid at 100°C for 8 h and subsequently was labeled with 1-phenyl-3-methyl-5-pyrazolone (PMP), and then the PMP-labeled monosaccharide were separated and identified by HPLC-UV at 250 nm as described in the experimental section. Peaks: 1. mannose, 2. ribose, 3. rhamnose, 4. glucuronic acid, 5. galacturonic acid, 6. glucose, 7. galactose, 8. arabinose, 9. fucose (internal standard).</p

PAP-3 induced apoptosis on MCF-7 cells.

<p>(A) MCF-7 cells were treated with 200 µg/mL PAP-3 for 12, 24, and 48 h, and the morphological changes were determined by fluorescence microscopy. (B) Cells were double-stained with annexin V-FITC and PI, and then cells were analyzed by flow cytometry. All experiments were done independently in triplicate per experimental point, and representative results are shown. The results represent the mean ± SD of three independent experiments. *<i>p</i><0.05 and **<i>p</i><0.01 indicate statistically significant differences versus control group.</p

Cell cycle analysis of PAP-3-treated cells.

<p>Cells were harvested and fixed in 70% alcohol and then stained with propidium iodide. Finally the stained cells were analyzed using a flow cytometer.</p

Fluoroscence data of DNA Neurons execute XOR logic with input “01”.

<p>The X-axis is cycling time of Real-time PCR, the time span of each cycle is 10 minutes, temperature of each cycle keeps in 24–25°C. The Y-axis is relative intensity of HEX (red curve) and FAM (blue curve).</p

Probes with Fluorophores.

<p>Probes with Fluorophores.</p

Fluoroscence data of DNA Neurons execute XOR logic with input “11”.

<p>The X-axis is cycling time of Real-time PCR, the time span of each cycle is 10 minutes, temperature of each cycle keeps in 24–25°C. The Y-axis is relative intensity of HEX (red curve) and FAM (blue curve).</p

Domains of “DNA neurons”.

<p>Domains of “DNA neurons”.</p

Abstract diagrams of two DNA neurons executing XOR logic.

<p>Abstract diagrams of two DNA neurons executing XOR logic.</p