Journal: Children

Article Title: Perspective on Similarities and Possible Overlaps of Congenital Disease Formation—Exemplified on a Case of Congenital Diaphragmatic Hernia and Neuroblastoma in a Neonate

doi: 10.3390/children8020163

Figure Lengend Snippet: H&E (Hematoxylin and Eosin) and immunohistochemical staining of primary tumor and metastases. ( A ) H&E staining of primary tumor shows histomorphological characteristics of poorly differentiated, Schwannian stroma poor neuroblastoma; ( B ) aquaporin 1 (AQP1) immunohistochemical staining and hematoxylin counterstaining of the liver metastases reveals dissolution of the periportal structure in the tumor-infiltrated regions. AQP1 positivity is observed in metastatic areas as well as in vascular endothelium (which can serve as an internal positive control). ( C ) AQP1, CXCR4 and MAP2 (microtubule associated protein) immunofluorescence staining of the primary tumor. Nuclei staining with DAPI (4′,6-diamidin-2-phenylindol). Next to AQP1, which is expressed in vascular structures as well as specific tumor areas, there is some expression of CXCR4 as a marker for metastatic homing as well as MAP2 expression in some areas indicating a more mature neuronal phenotype. Overall, tumor and metastases express experimental features that can be associated with a more aggressive phenotype.

Article Snippet: Immunohistochemistry staining was performed using the HRP-AEC-System from R&D Systems (Biotechnology Company, Minneapolis, MN, USA) with polyclonal rabbit anti-AQP1 antibody (Merck Millipore, Darmstast, Germany) at a dilution of 1:400 and counterstaining was performed with Mayer’s hematoxylin solution (Spitalpharmazie, Basel, Switzerland).

Techniques: Immunohistochemical staining, Staining, Positive Control, Immunofluorescence, Expressing, Marker

Journal: Cell & Bioscience

Article Title: Long non-coding RNA CASC2 improved acute lung injury by regulating miR-144-3p/AQP1 axis to reduce lung epithelial cell apoptosis

doi: 10.1186/s13578-018-0205-7

Figure Lengend Snippet: The expression level of CASC2 was decreased in ALI mice. The right lung tissues were respectively isolated from control mice and ALI mice. a The CASC2 expression was significantly decreased in ALI mice. b The miR-144-3p expression was significantly increased in ALI mice. c The AQP1 mRNA and protein expressions were both significantly decreased in ALI mice. *P < 0.05 vs control

Article Snippet: Primary mouse monoclonal antibodies against AQP1 and β-actin (Abcam, UK), and secondary antibody peroxidase-conjugated rabbit anti-IgG (Sigma) were used in western blot analysis.

Techniques: Expressing, Isolation, Control

Journal: Cell & Bioscience

Article Title: Long non-coding RNA CASC2 improved acute lung injury by regulating miR-144-3p/AQP1 axis to reduce lung epithelial cell apoptosis

doi: 10.1186/s13578-018-0205-7

Figure Lengend Snippet: The expression level of CASC2 was decreased in LPS-induced A549 cell. LPS-induced the Human Type II Alveolar Epithelial Cell Line (A549) be as the experimental model in vitro. a The CASC2 expression was significantly decreased in LPS-induced A549. b The miR-144-3p expression was significantly increased in LPS-induced A549. c The AQP1 mRNA and protein expressions were both significantly decreased in LPS-induced A549. *P < 0.05 vs control

Article Snippet: Primary mouse monoclonal antibodies against AQP1 and β-actin (Abcam, UK), and secondary antibody peroxidase-conjugated rabbit anti-IgG (Sigma) were used in western blot analysis.

Techniques: Expressing, In Vitro, Control

Journal: Cell & Bioscience

Article Title: Long non-coding RNA CASC2 improved acute lung injury by regulating miR-144-3p/AQP1 axis to reduce lung epithelial cell apoptosis

doi: 10.1186/s13578-018-0205-7

Figure Lengend Snippet: Overexpression of CASC2 inhibited LPS-induced A549 cell apoptosis. To investigate the effect of CASC2 on LPS-induced A549 cell apoptosis, the cell transfected with pcDNA-CASC2 and its control vector, respectively, and then induced by LPS. a The pcDNA-CASC2 transfection reversed the LPS-reduced CASC2 expression in A549 cell. b The pcDNA-CASC2 transfection reversed the LPS-increased miR-144-3p expression in A549 cell. c The pcDNA-CASC2 transfection reversed the LPS-reduced AQP1 expression in A549 cell. d The pcDNA-CASC2 transfection reversed the LPS-induced cell apoptosis. *P < 0.05 vs control, # P < 0.05 vs pcDNA

Article Snippet: Primary mouse monoclonal antibodies against AQP1 and β-actin (Abcam, UK), and secondary antibody peroxidase-conjugated rabbit anti-IgG (Sigma) were used in western blot analysis.

Techniques: Over Expression, Transfection, Control, Plasmid Preparation, Expressing

Journal: Cell & Bioscience

Article Title: Long non-coding RNA CASC2 improved acute lung injury by regulating miR-144-3p/AQP1 axis to reduce lung epithelial cell apoptosis

doi: 10.1186/s13578-018-0205-7

Figure Lengend Snippet: CASC2 controlled AQP1 expression by regulating miR-144-3p. a The AQP1 mRNA expression level was decreased in LPS-induced A549 cell. Under LPS induced environment, transfection of pcDNA-CASC2 increased AQP1 mRNA expression level, while co-transfection of pcDNA-CASC2 and miR-144-3p mimic reversed the increased AQP1 mRNA expression level. b The AQP1 protein expression level in A549 cell were in accord with the mRNA expression. *P < 0.05 vs pcDNA, # P < 0.05 vs pcDNA-CASC2 + pre-NC, & P < 0.05 vs pcDNA-CASC2 + miR-144-3p mimic

Article Snippet: Primary mouse monoclonal antibodies against AQP1 and β-actin (Abcam, UK), and secondary antibody peroxidase-conjugated rabbit anti-IgG (Sigma) were used in western blot analysis.

Techniques: Expressing, Transfection, Cotransfection

Journal: Cell & Bioscience

Article Title: Long non-coding RNA CASC2 improved acute lung injury by regulating miR-144-3p/AQP1 axis to reduce lung epithelial cell apoptosis

doi: 10.1186/s13578-018-0205-7

Figure Lengend Snippet: AQP1 was a target of miR-144-3p in A549 cell. a According to the predicted binding site of miR-144-3p in AQP1 transcript, we constructed a mutant plasmid of AQP1. b The miR-144-3p inhibitor enhanced the luciferase activity in WT-AQP1-transfected A549 cell, while have no impact in MUT-AQP1-transfected A549 cell. Meanwhile, the AQP1 mRNA and protein expression levels were significantly increased in miR-144-3p inhibitor-transfected A549 cells. c The miR-144-3p mimic reduced the luciferase activity in WT-AQP1-transfected A549 cell, while have no impact in MUT-AQP1-transfected A549 cell. Meanwhile, the AQP1 mRNA and protein expression levels were significantly decreased in miR-144-3p mimic-transfected A549 cells. *P < 0.05 vs pre-NC, # P < 0.05 vs NC

Article Snippet: Primary mouse monoclonal antibodies against AQP1 and β-actin (Abcam, UK), and secondary antibody peroxidase-conjugated rabbit anti-IgG (Sigma) were used in western blot analysis.

Techniques: Binding Assay, Construct, Mutagenesis, Plasmid Preparation, Luciferase, Activity Assay, Transfection, Expressing

Journal: Cell & Bioscience

Article Title: Long non-coding RNA CASC2 improved acute lung injury by regulating miR-144-3p/AQP1 axis to reduce lung epithelial cell apoptosis

doi: 10.1186/s13578-018-0205-7

Figure Lengend Snippet: LPS induced A549 cell apoptosis by regulating CASC2/miR-144-3p/AQP1 axis. The A549 cell was transfected or co-transfected of pcDNA-CASC2, miR-144-3p mimic and pcDNA-AQP1 and induced by LPS. And then the cell apoptosis was detected by MTT, and demonstrated by caspase-3 expression. a The inhibited cell apoptosis by pcDNA-CASC2 was reversed by miR-144-3p mimic, while which was reversed again by pcDNA-AQP1. b The caspase-3 expression by western blot. c A549 cell apoptosis in different groups using flow cytometry assay. *P < 0.05 vs pcDNA, # P < 0.05 vs pre-NC, & P < 0.05 vs miR-144-3p mimic

Article Snippet: Primary mouse monoclonal antibodies against AQP1 and β-actin (Abcam, UK), and secondary antibody peroxidase-conjugated rabbit anti-IgG (Sigma) were used in western blot analysis.

Techniques: Transfection, Expressing, Western Blot, Flow Cytometry

Journal: FEBS Open Bio

Article Title: Escin suppresses HMGB1‐induced overexpression of aquaporin‐1 and increased permeability in endothelial cells

doi: 10.1002/2211-5463.12622

Figure Lengend Snippet: AQP1 immunofluorescence changes after treated with HMGB1 or HMGB1 combined with escin in cultured ECs. (A) Normal AQP1 expression intensity in ECs. (B) Treatment with HMGB1 (100 ng·mL −1 ) for 24 h significantly increased AQP1 expression intensity and produced some deformity of ECs. (C) Treatment with HMGB1 (100 ng·mL −1 ) combined with escin (10 μg·mL −1 ) for 24 h led to a comparatively reduced AQP1 expression intensity. (D–F) AQP1 expression intensity was significantly decreased when ECs were treated with HMGB1 (100 ng·mL −1 ) combined with escin (20 μg·mL −1 ) for 12, 18, or 24 h. Scale bar = 200 μm.

Article Snippet: The coverglasses, on which the ECs grew, were removed and rinsed with PBS (Solarbio) at room temperature for 20 min. After being washed three times with PBS, cells were blocked with 5% BSA (Guangzhou Saiguo Biotech Co., Ltd, Guangzhou, China) in PBS for 30 min at room temperature, and subsequently incubated with primary antibody against AQP1 (1 : 200, Proteintech Group, Inc., Rosemont, IL, USA) at 4 °C overnight.

Techniques: Immunofluorescence, Cell Culture, Expressing, Produced

Journal: Scientific Reports

Article Title: Unilateral nephrectomy elongates primary cilia in the remaining kidney via reactive oxygen species

doi: 10.1038/srep22281

Figure Lengend Snippet: Mice were subjected either to UNx or a sham (control) operation. Cohorts of mice were treated with either MnTMPyP (5 mg/kg body weight) or saline (vehicle) daily, beginning on day 1 following surgery. Nine days following surgery, the remaining kidney was harvested. ( A – E ) To detect primary cilia, paraffin-embedded kidney sections were subjected to immunofluorescene staining using anti-acetylated α-tubulin antibody. AQP-1- and AQP-2-positive cell staining indicates the proximal tubule and collecting duct, respectively. AQP-1 and -2 negative cells were considered to be distal tubule cells. Primary cilia length was measured in the ( A , B ) parietal, ( A , C ) proximal tubule (PT), ( A , D ) distal tubule (DT), and ( A , E ) collecting duct (CD) cells. The length of 30 primary cilia per kidney were averaged (n = 4). Green color shows acetylated tubulin-positive primary cilia. Red color indicates AQP-1- ( B ) or AQP-2 ( D )-positive cells. DAPI (blue) stains nuclei. Results are expressed as the means ± SE. * p < 0.05 vs. respective-control., # p < 0.05 vs. UNx-vehicle on the same day.

Article Snippet: Sections were blocked with PBS containing 1% bovine serum albumin (blocking buffer) for 30 minutes and then incubated with anti-acetylated tubulin (Sigma-Aldrich), -AQP-1 (Alomone Labs) and -AQP-2 (Alomone Labs) antibodies diluted in blocking buffer overnight at 4 °C.

Techniques: Staining