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. 2017 Jan 27;7:41647. doi: 10.1038/srep41647

Figure 5. Effects of rhCygb on NADPH oxidase activation, ROS, O2 and NO generation in rat Kupffer cells.

Figure 5

(a) NADPH oxidase activity. Cultured KCs were treated with LPS (10 μg/mL) in the presence or absence of variant doses of rhCygb (0, 5, 10 or 20 μg/mL) for 24 h. NADPH oxidase activity was then measured. Data are expressed as mean ± S.D. (n = 6). ***P < 0.001 vs. normal control; ###P < 0.001 vs. LPS control; &P < 0.05 vs. 5, 10 μg/mL rhCygb. (b–e) Intracellular productions of ROS, O2 and NO. Intracellular ROS levels were assessed using a fluorescent probe, DCFH-DA. Fluorescence intensity was detected by multimode reader (b) and observed directly under the fluorescence microscope (e upper panels). The light green fluorescence represented intracellular ROS. Intracellular O2 was detected by a fluorescent probe, DHE. Fluorescence intensity was detected by multimode reader (c) and observed directly under the fluorescence microscope (e, middle panels). The red fluorescence represented intracellular O2. Intracellular NO was detected by a fluorescent probe, DAF-FM DA. Fluorescence intensity was detected by multimode reader (d) and observed directly under the fluorescence microscope (e, lower panels). The green fluorescence represented intracellular NO. Data are expressed as mean ± S.D. (n = 12). ***P < 0.001 vs. normal control; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. LPS control; &P < 0.05 vs. 5, 10 μg/mL rhCygb.

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