Journal: Oncology Letters
Article Title: Notch4 inhibition reduces migration and invasion and enhances sensitivity to docetaxel by inhibiting Akt/fascin in pancreatic cancer cells
Figure Lengend Snippet: Notch4 siRNA regulates the expression of p-Akt, Akt, GSK3, p-GSK3 and fascin in pancreatic cancer cells. The protein expression levels were determined by western blot analysis. Akt, p-Akt and fascin protein expression were markedly decreased in the Notch4 siRNA + docetaxel group compared with that in the Notch4 siRNA and docetaxel groups. GAPDH was used as an internal control to demonstrate equal protein loading. Lane 1, blank control; lane 2, control siRNA; lane 3, Notch4 siRNA; lane 4, docetaxel; lane 5, Notch4 siRNA + docetaxel. GSK3, glycogen synthase kinase 3; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; p, phosphorylated; siRNA, small interfering RNA.
Article Snippet: After blocking for 2 h with phosphate-buffered saline containing 0.1% Tween 20 (PBST) and 5% powdered skim milk, the blots were incubated with primary Notch4 (cat. no. ab134831; 1:1,000), p-Akt (cat. no. ab8933; 1:500), Akt (cat. no. ab8805; 1:1,000), GSK3 (cat. no. ab131344; 1:500), p-GSK3 (cat. no. ab28808; 1:500) and fascin (cat. no. ab97753; 1:1,000) antibodies (Abcam, Cambridge, UK) overnight in 5% powdered skim milk buffer, washed thrice with PBST and incubated with horseradish peroxidase-conjugated anti-rabbit (cat. no. 7074) and anti-mouse (cat. no. 7076) IgG secondary antibodies (1:2,000; Cell Signaling Technology, Beverly, MA, USA).
Techniques: Expressing, Western Blot, Small Interfering RNA