Journal: Oncology reports

Article Title: Activation of the FAK/PI3K pathway is crucial for AURKA-induced epithelial-mesenchymal transition in laryngeal cancer.

doi: 10.3892/or.2016.4872

Figure Lengend Snippet: Figure 3. Downregulation of AURKA using VX680 inhibits the phosphorylation of the FAK/PI3K pathway in Hep2 cells. (A) Effects of VX680 on the expression of related proteins were analyzed by western blotting. Expression of p-AURKA, p-FAK (Tyr397) and p-PI3K was reduced. (B) Quantification of the protein ratio (**P<0.01).

Article Snippet: Hep2 cells, treated with vX680, the inhibitor of FAK (TAE226), and the inhibitor of PI3K (omipalisib) (both from Selleck Chemicals), respectively, were lysed with RIPA buffer containing 1% protease inhibitor cocktail (100:1) and quantified by the BCA protein assay kit (all from Pierce, Rockford, IL, USA) according to the manufacturer's instructions.

Techniques: Phospho-proteomics, Expressing, Western Blot

Journal: Oncology reports

Article Title: Activation of the FAK/PI3K pathway is crucial for AURKA-induced epithelial-mesenchymal transition in laryngeal cancer.

doi: 10.3892/or.2016.4872

Figure Lengend Snippet: Figure 4. Effects of the inhibition of FAK/PI3K using TAE226 or omipalisib in Hep2 cells. (A) Effects of TAE226 on the expression of EMT-related proteins were analyzed by western blotting. The expression of E-cadherin was increased, whereas N-cadherin, vimentin and slug were decreased. (B) Quantification of EMT-related protein ratio after treatment with TAE226 (*P<0.05, **P<0.01). (C) Effects of TAE226 on the expression of p-AURKA, AURKA, p-FAK (Tyr397), FAK, p-PI3K and PI3K were analyzed by western blotting. Expression of p-FAK (Tyr397) and p-PI3K was decreased, while the others were not changed. (D) Quantification of the protein ratio after treatment with TAE226 (*P<0.05, **P<0.01). (E) Effects of omipalisib on the expression of EMT-related proteins as analyzed by western blotting. Expression of E-cadherin was increased, whereas N-cadherin, vimentin and slug were decreased. (F) Quantification of EMT- related protein ratio after treatment with omipalisib (*P<0.05, **P<0.01). (G) Effects of omipalisib on the expression of the regulatory factors as analyzed by western blotting. Only p-PI3K was decreased. (H) Quantification of related protein ratio after treatment with omipalisib (**P<0.01).

Article Snippet: Hep2 cells, treated with vX680, the inhibitor of FAK (TAE226), and the inhibitor of PI3K (omipalisib) (both from Selleck Chemicals), respectively, were lysed with RIPA buffer containing 1% protease inhibitor cocktail (100:1) and quantified by the BCA protein assay kit (all from Pierce, Rockford, IL, USA) according to the manufacturer's instructions.

Techniques: Inhibition, Expressing, Western Blot

Journal: Oncology reports

Article Title: Activation of the FAK/PI3K pathway is crucial for AURKA-induced epithelial-mesenchymal transition in laryngeal cancer.

doi: 10.3892/or.2016.4872

Figure Lengend Snippet: Figure 5. Inhibition of the FAK/PI3K pathway by TAE226/omipalisib reduces mobility, migration and invasion in Hep2 cells. (A) Effects of TAE226/omipal- isib on cell mobility in the wound healing assay. FAK/PI3K promoted Hep2 cell mobility. (B) Quantification of wound size (*P<0.05). (C) Effects of TAE226/ omipalisib on cell migration and invasion by migration and invasion assays. FAK/PI3K promoted the migration and invasion of Hep2 cells. (D) Quantification of cell number/field (*P<0.05, **P<0.01).

Article Snippet: Hep2 cells, treated with vX680, the inhibitor of FAK (TAE226), and the inhibitor of PI3K (omipalisib) (both from Selleck Chemicals), respectively, were lysed with RIPA buffer containing 1% protease inhibitor cocktail (100:1) and quantified by the BCA protein assay kit (all from Pierce, Rockford, IL, USA) according to the manufacturer's instructions.

Techniques: Inhibition, Migration, Wound Healing Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: MFG-E8 Maintains Cellular Homeostasis by Suppressing Endoplasmic Reticulum Stress in Pancreatic Exocrine Acinar Cells

doi: 10.3389/fcell.2021.803876

Figure Lengend Snippet: rhMFG-E8 alleviates ER stress in pancreatic exocrine acinar cells through integrin αVβ3/5-FAK-STAT3. Pancreatic AR42J cells (5 × 10 5 /well) were treated with 100 nmol/L cerulein and 10 ng/ml LPS with or without 100 ng/ml MFG-E8 for 24 h. To determine whether the protective effect of MFG-E8 in ER stress is mediated through integrin αVβ3/5-FAK-STAT3, 5 ng/ml cilengitide, a specific integrin αVβ3/5 antagonist, 1 μmol/L-PF-00562271, a specific FAK antagonist, 30 μmol/L-APTSTAT3-9R, a specific STAT3 antagonist, was administered with 100 ng/ml MFG-E8, respectively. (A,B) Western blot analysis of the expression of GRP78, phospho-IRE1α, IRE1α, CHOP, caspase-9 and cleaved caspase-9 in the AR42J cells; (C–H) Western blot analysis of the expression of phospho-FAK, FAK, phospho-STAT3 and STAT3 in the AR42J cells; (I,J) Western blot analysis of the expression of GRP78, phospho-IRE1α, IRE1α, CHOP, caspase-9 and cleaved caspase-9 in the AR42J cells. n = 6/group, error bars indicate the SEM; ∗ p < 0.05 versus Sham group; # p < 0.05 versus Vehicle group. MFG-E8, milk fat globule EGF factor 8; GRP78, glucose-regulated protein 78; CHOP, C/EBP homologous protein; LPS, Lipopolysaccharide.

Article Snippet: The AR42J cells were treated with 100 nmol/L cerulein (C6660, Solarbio, United States) and 10 ng/ml Lipopolysaccharide (LPS) (L-8880, Solarbio, United States) with or without 20 or 100 ng/ml MFG-E8 (2805-MF, RD System, Inc. Minnesota, United States) for 24 h. In additional groups of AR42J cells, Cilengitide (5 ng/ml, SELLECK, Texas, United States), a specific αvβ3/5 integrin inhibitor or PF00562271 (S2672, SELLECK, Texas, United States), a specific FAK inhibitor or APTSTAT3-9R (S8197, SELLECK, Texas, United States), a specific STAT3 antagonist, were administered together with 100 ng/ml MFG-E8.

Techniques: Western Blot, Expressing