phosphorylated erk af1018  (R&D Systems)

Journal: Oncology Letters

Article Title: Rab23 contributes to the progression of colorectal cancer via protein kinase B and extracellular signal-regulated kinase signaling pathways

doi: 10.3892/ol.2019.10491

Figure Lengend Snippet: ERK and AKT signaling pathways are required for regulatory effect of Rab23 on colorectal cancer cell proliferation. Rab23 overexpression in (A) SW1116 and (B) HT29 cells resulted in increased phosphorylation of ERK and AKT. The expression of Ki-67 in (C) SW1116 and (D) HT29 cells was inhibited by addition of the ERK inhibitor U0126 or AKT inhibitor LY294002. (E) SW1116 and (F) HT29 cell proliferation ability was inhibited by addition of the ERK inhibitor U0126 or AKT inhibitor LY294002. *P<0.05 vs. Vector-Rab23. ERK, extracellular signal-regulated kinase; AKT, protein kinase B; CTL, control; OD, optical density; LY, AKT inhibitor LY294002.

Article Snippet: Subsequent to blocking in 5% non-fat milk (cat. no. 232100; Becton, Dickinson and Company, New Jersey, USA) at room temperature for 1 h, the membrane was probed at 4°C overnight using the following primary antibodies: ERK (cat. no. AF1576; 1:1,000), phosphorylated (p)-ERK (cat. no. AF1018; 1:1,000), AKT (cat. no. AF2055; 1:1,000) and p-AKT (cat. no. AF887; 1:1,000), purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA); Rab23 (cat. no. ab230200; 1:300), Ki-67 (cat. no. ab833; 1:300) and GAPDH (cat. no. ab9485; 1:4,000), all purchased from Abcam (Cambridge, MA, USA).

Techniques: Over Expression, Expressing, Plasmid Preparation