Resultative Compound Verb in Modern Chinese : A Comment on Imai(1985) and Lu(1986)

<p>A. API and DMO suppresses NF-κB DNA binding ability in HCT116 cells. HCT116 cells were treated with DMO and API at indicated doses, nuclear extracts were prepared, and 20 μg of the nuclear extract protein was used for the ELISA-based DNA-binding assay *p<0.05; **p<0.005). B & C. NF-κB responsive elements linked to a luciferase reporter gene were transfected with wild-type or dominant-negative IκB and transfected cancer cells were treated at indicated doses for 6 h and luciferase activity was measured as described in Materials and Methods section. All luciferase experiments were done in triplicate and repeated twice (*p<0.05; **p<0.005). D. API abrogates constitutive IκBα phosphorylation in dose-dependent manner in HCT116 cells. HCT116 cells were treated with different concentrations of API (0, 5, 10 and 20 μM) for 6 h and cytoplasmic extract was prepared. Lysates were resolved on SDS gel and electrotransferred to a nitrocellulose membrane and probed with anti-phospho-IκBα/IκBα. The blot was washed, exposed to HRP-conjugated secondary antibodies for 1 h, and finally examined by chemiluminescence. GAPDH was used as loading control.</p

In silico interaction between the oxazines and IκBα/NF-κB complex.

<p>A. Representation of the native IκBα/NF-κB heterodimer and docked solution of tested oxazines (in stick representation). The sub-unit of p50 is represented in green, p65 in cyan, and IκBα in pink. B. Molecular docking of the lead structure API with the NF-κB heterodimer solution was shown. C. Interaction map lead compound API that bound with key amino acids of the crystal structure. Hydrogen bonding (black dots) between API oxygen atoms with Tyr251, and other Asp271, Arg246, His245 was shown.</p

Novel Synthetic Oxazines Target NF-κB in Colon Cancer <i>In Vitro</i> and Inflammatory Bowel Disease <i>In Vivo</i> - Fig 1

<p>A. The lead compounds identified among the 1,2-oxazine derivatives tested decreased the cell proliferation of HCT116 cells in dose-dependent manner. B. Microscopic images to demonstrate the inhibition of cell proliferation (Magnification 4x). C. HCT116 cells were treated with API for 48 h and caspase 3/7 assay was performed. We observed a significant dose dependent increase in caspase 3/7 activity levels demonstrating the ability of API to induce apoptosis.</p

Novel Synthetic Oxazines Target NF-κB in Colon Cancer <i>In Vitro</i> and Inflammatory Bowel Disease <i>In Vivo</i> - Fig 3

<p>A. API and DMO suppresses NF-κB DNA binding ability in HCT116 cells. HCT116 cells were treated with DMO and API at indicated doses, nuclear extracts were prepared, and 20 μg of the nuclear extract protein was used for the ELISA-based DNA-binding assay *p<0.05; **p<0.005). B & C. NF-κB responsive elements linked to a luciferase reporter gene were transfected with wild-type or dominant-negative IκB and transfected cancer cells were treated at indicated doses for 6 h and luciferase activity was measured as described in Materials and Methods section. All luciferase experiments were done in triplicate and repeated twice (*p<0.05; **p<0.005). D. API abrogates constitutive IκBα phosphorylation in dose-dependent manner in HCT116 cells. HCT116 cells were treated with different concentrations of API (0, 5, 10 and 20 μM) for 6 h and cytoplasmic extract was prepared. Lysates were resolved on SDS gel and electrotransferred to a nitrocellulose membrane and probed with anti-phospho-IκBα/IκBα. The blot was washed, exposed to HRP-conjugated secondary antibodies for 1 h, and finally examined by chemiluminescence. GAPDH was used as loading control.</p

Serum cytokine profile of control and experimental mice were estimated using murine mini ELISA development kits according to the manufacturer’s protocol.

<p>A. TNF-α. B. IFN-γ. C. IL-1β. D. IL-6. and E. IL-10. SZ-Sulfasalazine. Data are presented as mean ± S.E.M. * <i>p</i><0.05; **<i>p</i>< 0.01; ***<i>p</i><0.001.</p