Journal: International Journal of Molecular Sciences

Article Title: Evodiamine Induces Apoptosis in SMMC-7721 and HepG2 Cells by Suppressing NOD1 Signal Pathway

doi: 10.3390/ijms19113419

Figure Lengend Snippet: Evo decreases expression of NOD1 resulting in suppression of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) activation in vitro. ( A ) Expression of NOD1 in normal hepatocyte HL-7702, HepG2, and SMMC-7721 cells were detected by qRT-PCR and Western blot assays. ( B ) HepG2 and SMMC-7721 cells were incubated with 1 µM Evo for 0, 3, 6, and 12 h. The Western blot assay was performed to detect levels of proteins in the NOD1 pathway. ( C ) Representative images from the immunofluorescence method (×400). HepG2 and SMMC-7721 cells were incubated with or without Evo (0, 0.5, and 1 µM) for 24 h to detect levels of NOD1 and p-P65. Values are means and standard errors of three separate experiments (* p < 0.05 and ** p < 0.01 versus control).

Article Snippet: Antibody against NOD1 (NB100-56152) was purchased from Novus Biologicals (San Diego, CA, USA).

Techniques: Expressing, Activation Assay, In Vitro, Quantitative RT-PCR, Western Blot, Incubation, Immunofluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Evodiamine Induces Apoptosis in SMMC-7721 and HepG2 Cells by Suppressing NOD1 Signal Pathway

doi: 10.3390/ijms19113419

Figure Lengend Snippet: Evo induces apoptotic cell death of HepG2 and SMMC-7721 cells via the NOD1-mediated apoptotic pathway in vitro. HepG2 and SMMC-7721 cells were treated with 10 µg/mL IE-DAP for 2 h before exposure to 1 µM Evo. Apoptosis ( A ), cycle arrest ( B ), apoptosis-related protein levels ( C ) and the proteins’ levels of NOD1 pathway ( D ) in treated and untreated cells were measured by 5-ethynyl-2′-deoxyuridine (EdU) (×40), cellular propidium iodide (PI) fluorescence and Western blot methods. The percentage of proliferating cells (EdU+) was quantitated using ImageJ software (National Institutes of Health, Bethesda, MD, USA). Values are means and standard errors of three separate experiments (* p < 0.05 and ** p < 0.01 versus control, # p < 0.05 and ## p < 0.01 versus Evo).

Article Snippet: Antibody against NOD1 (NB100-56152) was purchased from Novus Biologicals (San Diego, CA, USA).

Techniques: In Vitro, Fluorescence, Western Blot, Software, Control

Journal: International Journal of Molecular Sciences

Article Title: Evodiamine Induces Apoptosis in SMMC-7721 and HepG2 Cells by Suppressing NOD1 Signal Pathway

doi: 10.3390/ijms19113419

Figure Lengend Snippet: Evo-induced apoptosis of HCC cells occurred via the NOD1 pathway in vivo. ( A , B ) The mice treated with or without 10 mg/kg of Evo. Levels of proteins in the NOD1 pathway in tumor tissues were detected by the Western blot method. ( C ) Representative images from the immunofluorescence method. Expression of NOD1 (×400) and p-P65 (×200) in tumor tissues was analyzed by the immunofluorescence method. Values are means and standard errors of five separate experiments (* p < 0.05 and ** p < 0.01 versus control).

Article Snippet: Antibody against NOD1 (NB100-56152) was purchased from Novus Biologicals (San Diego, CA, USA).

Techniques: In Vivo, Western Blot, Immunofluorescence, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Evodiamine Induces Apoptosis in SMMC-7721 and HepG2 Cells by Suppressing NOD1 Signal Pathway

doi: 10.3390/ijms19113419

Figure Lengend Snippet: Model of NOD1 signaling cascade. Nucleotide-binding oligomerization domain 1 (NOD1) characterized by three motifs containing a C-terminal leucine-rich repeat (LRR) domain, a central nucleotide-binding domain (NBD), and a variable N-terminal caspase recruitment domains (CARD), which could be activated by γ-D-Glu-mDAP (IE-DAP). NOD1 recruits the adaptor RIP2 and facilitates recruitment of TAK1 and NEMO. Here the TAK1 recruits TAB1 and TAB2/3 inducing MAPK activation. In addition, NEMO instigates activation of NF-κB by reducing IκBα and phosphorylating P65, the subunit of NF-κB. Furthermore, activation of MAPK and NF-κB promotes the cell cycle and proliferation. In our study, Evodiamine could potentially suppress the NOD1 signal pathway to inhibit the cell cycle and proliferation. The arrows represents the activation and the T bar represents the suppression.

Article Snippet: Antibody against NOD1 (NB100-56152) was purchased from Novus Biologicals (San Diego, CA, USA).

Techniques: Binding Assay, Activation Assay