polyclonal primary antibodies for aqp1  (Boster Bio)

Journal: Particle and Fibre Toxicology

Article Title: Inhaled gold nanoparticles cause cerebral edema and upregulate endothelial aquaporin 1 expression, involving caveolin 1 dependent repression of extracellular regulated protein kinase activity

doi: 10.1186/s12989-019-0324-2

Figure Lengend Snippet: Au-NPs induced aquaporin-1 (AQP1) protein expression in bEnd.3 cells. a / b The bEnd.3 cells (an immortalized mouse cerebral endothelial cell line) were exposed to Au-NPs (or Au-MPs) and the expression level of AQP1 was detected by western blots. Representative images showed an increase of AQP1 protein level in Au-NP-treated groups, whereas AQP1 protein level remained unaffected in Au-MP-treated groups. a concentration-dependent treatment; cells were incubated with 10, 50, 100 and 500 ng/mL Au-NPs for 24 h. b time-dependent treatment; cells were incubated with 500 ng/mL Au-NPs for 3, 6, 12, and 24 h. (* p < 0.05, ** p < 0.01, and *** p < 0.001 indicates statistically significant difference from the control group; N = 11). c Representative images of immunofluorescent staining, the Au-NP-induced AQP1 and the nucleus was manifested by red and blue fluorescence, respectively. A gain of red fluorescence in cell membrane and cytosol was observed in Au-NP-treated bEnd.3 cells (500 ng/mL; 24 h), as compared to control. d Transendothelial permeability assay was performed as described in Materials and Methods. Au-NP treatment (500 ng/mL; 24 h) made bEnd.3 cell more permeable to water. (* p < 0.05, indicates statistically significant difference from the control group; N = 12)

Article Snippet: After Au-NP treatment (500 ng/mL; 24 h), the cells were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 for 5 min at room temperature, and incubated with blocking serum for 30 min. Next, the cells were incubated overnight (4 °C) with a primary AQP1 antibody (1:50; Bioss Antibodies, Woburn, MA, catalog. bs-1506R) and then with a fluorescein isothiocyanate (FITC)-conjugated secondary antibody (1:1000) at room temperature for 1 h in the dark.

Techniques: Expressing, Western Blot, Concentration Assay, Incubation, Staining, Fluorescence, Permeability

Journal: Particle and Fibre Toxicology

Article Title: Inhaled gold nanoparticles cause cerebral edema and upregulate endothelial aquaporin 1 expression, involving caveolin 1 dependent repression of extracellular regulated protein kinase activity

doi: 10.1186/s12989-019-0324-2

Figure Lengend Snippet: Au-NP treatment rapidly changed the phosphorylating status of FAK, AKT and ERK, and subsequently contributing to Au-NP-mediated AQP1 accumulation. bEnd.3 cells were incubated with 500 ng/mL Au-NPs for 15, 30 and 60 min, and the phosphorylating status of FAK, AKT, ERK, and Cav1 was measured by western blots. a Representative images showed an augmentation of FAK and AKT phosphorylation; a reduction of ERK and Cav1 phosphorylation in Au-NP-treated groups in time-dependent manner, as compared to control. Also, Au-NP treatment caused an accumulation of Cav1 protein level. Quantified data was gained by densitometry analysis, followed by a normalized process to their total form. b phospho-FAK, ( c ) phospho-AKT, ( d ) phospho-ERK, ( e ) Cav1 and ( f ) phospho-Cav1. (* p < 0.05, ** p < 0.01, and *** p < 0.001 indicates statistically significant difference from the control group; N > 10). g Cells were pre-incubated with 10 μM U0126 (ERK inhibitor), 10 μM GDC-0068 (pan-AKT inhibitor) and 10 μM PF-573228 (FAK inhibitor), subjected to a 12–16 h exposure of Au-NPs. Images and quantified data revealed that Au-NP-induced AQP1 expression was prevented while FAK and AKT inhibition, whereas an enhancement of AQP1 expression was presented in the presence of U0126. (* p < 0.05, ** p < 0.01, and *** p < 0.001 indicates statistically significant difference from the control group; ## p < 0.01, and ### p < 0.0001 indicates statistically significant difference from the Au-NP-treated group; N = 7)

Article Snippet: After Au-NP treatment (500 ng/mL; 24 h), the cells were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 for 5 min at room temperature, and incubated with blocking serum for 30 min. Next, the cells were incubated overnight (4 °C) with a primary AQP1 antibody (1:50; Bioss Antibodies, Woburn, MA, catalog. bs-1506R) and then with a fluorescein isothiocyanate (FITC)-conjugated secondary antibody (1:1000) at room temperature for 1 h in the dark.

Techniques: Incubation, Western Blot, Expressing, Inhibition

Journal: Particle and Fibre Toxicology

Article Title: Inhaled gold nanoparticles cause cerebral edema and upregulate endothelial aquaporin 1 expression, involving caveolin 1 dependent repression of extracellular regulated protein kinase activity

doi: 10.1186/s12989-019-0324-2

Figure Lengend Snippet: Caveolin-1 (Cav1) is a crucial factor in Au-NP-mediated AQP1 expression. a bEnd.3 cells with a silenced Cav1 (bEnd.3 Cav1-KD) was established as described in Materials and Methods. The mRNA and protein expression level of Cav1 and AQP1 was obviously down-regulated. b / c Both images and quantitative data proved Au-NP-mediated FAK and AKT phosphorylation (as well as ERK de-phosphorylation) were prevented in bEnd.3 Cav1-KD ( N = 6). d Au-NP-mediated AQP1 induction and ( e ) water permeability was also ameliorated in bEnd.3 Cav1-KD, as compared to wild-type (N = 6). These data manifested the requirement of Cav1 of bEnd.3 cell in respond to Au-NP stimulation. (* p < 0.05, ** p < 0.01, and *** p < 0.001 indicates statistically significant difference from the control group; ## p < 0.01, and ### p < 0.001 indicates statistically significant difference from the Au-NP-treated group)

Article Snippet: After Au-NP treatment (500 ng/mL; 24 h), the cells were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 for 5 min at room temperature, and incubated with blocking serum for 30 min. Next, the cells were incubated overnight (4 °C) with a primary AQP1 antibody (1:50; Bioss Antibodies, Woburn, MA, catalog. bs-1506R) and then with a fluorescein isothiocyanate (FITC)-conjugated secondary antibody (1:1000) at room temperature for 1 h in the dark.

Techniques: Expressing, De-Phosphorylation Assay, Permeability

Journal: Particle and Fibre Toxicology

Article Title: Inhaled gold nanoparticles cause cerebral edema and upregulate endothelial aquaporin 1 expression, involving caveolin 1 dependent repression of extracellular regulated protein kinase activity

doi: 10.1186/s12989-019-0324-2

Figure Lengend Snippet: Inhaled Au-NPs upregulated AQP1 expression in cerebral microvessels in vivo. a Representative images showed an increase in immunoreactivity against AQP1 in microvessels of brain tissues, whereas the stained CD31 (an endothelial biomarker) was without significant differences. b The quantitative data was determined by the measurement of the length of AQP1-positive microvessels. (** p < 0.01, and *** p < 0.001 indicates statistically significant difference from the control group, N = 4)

Article Snippet: After Au-NP treatment (500 ng/mL; 24 h), the cells were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 for 5 min at room temperature, and incubated with blocking serum for 30 min. Next, the cells were incubated overnight (4 °C) with a primary AQP1 antibody (1:50; Bioss Antibodies, Woburn, MA, catalog. bs-1506R) and then with a fluorescein isothiocyanate (FITC)-conjugated secondary antibody (1:1000) at room temperature for 1 h in the dark.

Techniques: Expressing, In Vivo, Staining, Biomarker Assay

Journal: International Journal of Molecular Sciences

Article Title: Personalized Treatment Response Assessment for Rare Childhood Tumors Using Microcalorimetry–Exemplified by Use of Carbonic Anhydrase IX and Aquaporin 1 Inhibitors

doi: 10.3390/ijms20204984

Figure Lengend Snippet: Clinical presentation ( A ) The child presented with a large abdominal mass, which was not reduced in size after initial chemotherapy. The scar was a result of the initial surgical biopsy. ( B ) MRI of primary clear cell sarcoma of left kidney following chemotherapy. Despite few necrotic areas the tumor is largely intact and appears to be vital. ( C ) Large tumor after resection. Immuno-staining ( D AQP1 IHC, E : CAIX IHC, F : AQP1 and CAIX IF stainings) The response to the specific inhibitors corresponded with the protein expression profiles of AQP1 (mostly in vibrovascular septae and vessels and CAIX (most strongly within tumor nests). Microcalorimteric measurements ( G ) Microcalorimetric measurements showed a decrease metabolic activity under treatment with AQP and CAIX inhibitors compared to medium control. (Control – medium, Drug 1—AQP1 inhibitor TEA, Drug 2—FC8-325A, Drug 3—FC8-207A, Drug 4—FC12-520A).

Article Snippet: AQP1 and CAIX staining was performed according to the protocol using a primary polyclonal rabbit anti-AQP1 antibody (Merck Millipore, Sigma-Aldrich Chemie GmbH Buchs, Switzerland) at a dilution of 1:400 and antibody M75 (BioScience Slovakia, Bratislava, Slovak republic) at a dilution of 1:200.

Techniques: Immunostaining, Expressing, Activity Assay