Journal: Journal of molecular medicine (Berlin, Germany)

Article Title: Acid sphingomyelinase inhibition protects mice from lung edema and lethal Staphylococcus aureus sepsis.

doi: 10.1007/s00109-014-1246-y

Figure Lengend Snippet: Fig. 4 Endothelial cells were infected for 2 h with S. aureus (MOI 10:1) or left uninfected. As indicated, cells were pretreated for 20 min with amitriptyline (Ami) (20 μM), Tiron (10 mM), or NAC (10 mM) before infection with S. aureus. Immunofluores- cence stainings were performed with antibodies against ZO1, ZO2, occludin, or E-cadherin for determination of the degradation of these TJ proteins. The present- ed pictures are representative of the results of at least three inde- pendent experiments. Scale bar is 25 μm

Article Snippet: Samples were washed and incubated overnight at 4 °C with antibodies against ZO1 (Invitrogen 40-2300, rabbit IgG), ZO2 (Santa Cruz Biotechnology Inc. sc-11448, rabbit IgG), occludin (71-1500, rabbit IgG, Invitrogen), or E-cadherin (Santa Cruz Biotechnology Inc. sc-7870, rabbit IgG).

Techniques: Infection

Journal: Cancer Management and Research

Article Title: ZNF259 promotes breast cancer cells invasion and migration via ERK/GSK3β/snail signaling

doi: 10.2147/CMAR.S174745

Figure Lengend Snippet: ZNF259 knockdown could downregulate phosphorylated-extracellular signal-regulated kinase (p-ERK), phosphorylated-glycogen synthase kinase 3β (p-GSK3β), and Snail expression, and upregulate E-cadherin and zonula occludens protein 1 (ZO-1) expression. Notes: Western blotting was performed after ZNF259 knockdown in both MCF-7 and MDA-MB-231 cell lines, and phosphorylated mitogen-activated protein kinase (p-MEK), phosphorylated-extracellular signal-regulated kinase (p-ERK), phosphorylated-glycogen synthase kinase 3β (p-GSK3β) (Ser 9), and Snail expressions were downregulated, while E-cadherin and ZO-1 expression was upregulated. The total MEK, ERK, and GSK3β levels did not show obvious changes. The numbers indicated the relative expression ratios of the protein. At least 3 separate experiments were performed. The statistical analysis histogram is shown in the . Abbreviation: ZNF259, zinc finger protein 259.

Article Snippet: Membranes were incubated overnight at 4°C with the following primary antibodies: ZNF259 (rabbit monoclonal antibody, 1:200, ab134970; Abcam; mouse monoclonal antibody, 1:200, sc-398491; Santa Cruz Biotechnology), phosphorylated mitogen-activated protein kinase (p-MEK), MEK, p-ERK, ERK, p-GSK3β, GSK3β, and Snail (1:500; Cell Signaling Technology, Danvers, MA, USA), E-cadherin (1:500; BD Transduction Laboratories, Lexington, KY, USA), zonula occludens protein 1 (ZO-1) (1:500; Proteintech, Chicago, IL, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:5,000; Sigma-Aldrich).

Techniques: Knockdown, Expressing, Western Blot

Journal: Cancer Management and Research

Article Title: ZNF259 promotes breast cancer cells invasion and migration via ERK/GSK3β/snail signaling

doi: 10.2147/CMAR.S174745

Figure Lengend Snippet: Invasion and migration promoted by ZNF259 were inhibited by the extracellular signal-regulated kinase (ERK) inhibitor U0126. Notes: ( A ) ZNF259 transfection could upregulate p-ERK, phosphorylated-glycogen synthase kinase 3β (p-GSK3β), and Snail expression, and downregulate E-cadherin and zonula occludens protein 1 (ZO-1) expression. All these effects could be restored by the incorporation of the ERK inhibitor U0126 (5 µM). The numbers indicated the relative expression ratios of protein. At least 3 separate experiments were performed. (B, C) Transwell assay showed that U0126 could reverse the ZNF259 transfection-caused increase in cell invasion ability (mean ± standard error of the mean, ** P <0.01). (D, E) Wound healing assay also showed that U0126 could reverse the ZNF259 transfection-caused increase in cell migration ability (mean ± standard error of the mean, * P <0.05). Abbreviation: ZNF259, zinc finger protein 259.

Article Snippet: Membranes were incubated overnight at 4°C with the following primary antibodies: ZNF259 (rabbit monoclonal antibody, 1:200, ab134970; Abcam; mouse monoclonal antibody, 1:200, sc-398491; Santa Cruz Biotechnology), phosphorylated mitogen-activated protein kinase (p-MEK), MEK, p-ERK, ERK, p-GSK3β, GSK3β, and Snail (1:500; Cell Signaling Technology, Danvers, MA, USA), E-cadherin (1:500; BD Transduction Laboratories, Lexington, KY, USA), zonula occludens protein 1 (ZO-1) (1:500; Proteintech, Chicago, IL, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:5,000; Sigma-Aldrich).

Techniques: Migration, Transfection, Expressing, Transwell Assay, Wound Healing Assay