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. 2019 Dec 18;25(1):13. doi: 10.3390/molecules25010013

Figure 3.

Figure 3

Figure 3

(A) Effect of MCA-1 on iNOS expression. J774 macrophages treated with 30 µg/mL of LPS to induce iNOS expression and total RNA were extracted to determine iNOS expression using RT-PCR, as described in the text. (B) Effect of MCA-1 on iNOS Protein (C)Effect of MCA-1 on NFkB phosphorylation. J774 macrophages were treated with LPS in the presence of varying concentrations of MCA-1 for 1 h to induce NFκB phosphorylation and for 48 h to monitor iNOS expression. A total of 35 μg of protein from the cell lysate was resolved on an SDS-PAGE and proteins were detected using specific antibodies against phosphorylated NFkB or iNOS, as described in the text. Sample loading was monitored by detecting β-actin with specific antibodies. The blots shown are representatives of two independent experiments. (D) The 3D structure of murine iNOS and the binding mode of MCA-1 at the active site of iNOS is shown in the box. The active site residues are depicted by the coral sticks, the active site of heme is shown by the purple sticks, and the compound is shown by the green sticks.