Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Effects of Cymbidium Root Ethanol Extract on Atopic Dermatitis

doi: 10.1155/2016/5362475

Figure Lengend Snippet: Effect of Cymbidium ethanol extract (CYM) on immunoglobulin E/dinitrophenyl-conjugated bovine serum albumin- (IgE/DNP-BSA-) induced phosphorylation of spleen tyrosine kinase (Syk), phospholipase C- (PLC-) γ , Akt, extracellular signal-regulated kinase (Erk) 1/2, c-Jun N-terminal kinase (JNK), and p38. RBL-2H3 cells were seeded and incubated in 24-well plates overnight in Minimum Essential Medium (MEM). Anti-DNP IgE (400 ng/mL) was added for 12 h, followed by addition of CYM (0 or 200 μ g/mL). Cells were then treated with DNP-BSA. Cells were lysed and protein was extracted by the PRO-PREP protein extraction kit. The protein lysates were analyzed by SDS-PAGE. (a) Western blot analysis for phosphorylation of Syk, PLC- γ , Akt, Erk1/2, JNK, and p38. (b) Densitometric analysis of expression of phosphorylated proteins. Data are mean ± standard deviation (SD, n = 3); ∗∗ P < 0.001 and ∗∗∗ P < 0.0001 versus sensitized RBL-2H3 cell.

Article Snippet: The primary antibodies used were anti-phospho-Syk, anti-Syk, anti-phospho-PLC- γ , anti-PLC- γ , anti-phospho-Akt, anti-Akt, anti-phospho-extracellular signal-regulated kinase (Erk) 1/2, anti-Erk 1/2, anti-phospho-c-Jun N-terminal kinase (JNK), anti-JNK, anti-phospho-p38, and anti-p38, which were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Incubation, Protein Extraction, SDS Page, Western Blot, Expressing, Standard Deviation