Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: CXCL12-induced directional migration requires AQP3 and H 2 O 2 uptake. (A to F) MDA-MB-231 and DU4475 cells were transfected with control or AQP3 siRNA. The error bars indicate SE. (A) The chemotaxis efficiency of control or AQP3 KD MDA-MB-231 and DU4475

Article Snippet: For AQP3 staining and visualization of macrophage infiltration, frozen or paraffin-embedded sections were stained with antibodies against AQP3 (Millipore), human EpCAM (eFluor 660; eBioscience), turbo-GFP (Origene), or F4/80 (allophycocyanin [APC]; Biolegend).

Techniques: Migration, Multiple Displacement Amplification, Transfection, Chemotaxis Assay

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: AQP3-dependent PTEN/PTP1B oxidation and Akt phosphorylation. (A to E) MDA-MB-231 cells were infected with lentiviral control or AQP3 shRNA, and DU4475 cells were transfected with control or AQP3 siRNA. (A) Immunoblot analysis of PTEN oxidation. Control

Article Snippet: For AQP3 staining and visualization of macrophage infiltration, frozen or paraffin-embedded sections were stained with antibodies against AQP3 (Millipore), human EpCAM (eFluor 660; eBioscience), turbo-GFP (Origene), or F4/80 (allophycocyanin [APC]; Biolegend).

Techniques: Multiple Displacement Amplification, Infection, shRNA, Transfection

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: Involvement of AQP3 expression in breast cancer cell migration in vivo . The error bars indicate SE. (A to D) MDA-MB-231 cells (8.0 × 10 5 cells) were intravenously injected into SCID mice, and the numbers of cells in the lungs after 24 h were analyzed

Article Snippet: For AQP3 staining and visualization of macrophage infiltration, frozen or paraffin-embedded sections were stained with antibodies against AQP3 (Millipore), human EpCAM (eFluor 660; eBioscience), turbo-GFP (Origene), or F4/80 (allophycocyanin [APC]; Biolegend).

Techniques: Expressing, Migration, In Vivo, Multiple Displacement Amplification, Injection, Mouse Assay

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: CXCL12-induced H 2 O 2 regulates PTP1B/PTEN oxidation and Akt activation. (A to C, E, and F) MDA-MB-231 cells were infected with lentiviral control or AQP3 shRNA. (A and B) The effects of DPI (20 μM; 30 min) or catalase (2,000 U/ml) on PTEN (A) or

Article Snippet: For AQP3 staining and visualization of macrophage infiltration, frozen or paraffin-embedded sections were stained with antibodies against AQP3 (Millipore), human EpCAM (eFluor 660; eBioscience), turbo-GFP (Origene), or F4/80 (allophycocyanin [APC]; Biolegend).

Techniques: Activation Assay, Multiple Displacement Amplification, Infection, shRNA

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: Transport of CXCL12-induced H 2 O 2 into breast cancer cells through AQP3. (A) The mRNA levels of AQP3 in control (si-control)- or AQP3-siRNA (si-AQP3)-transfected MDA-MB-231 (top) and DU4475 (bottom) cells and in lentiviral control (sh-control)- or AQP3-shRNA

Article Snippet: For AQP3 staining and visualization of macrophage infiltration, frozen or paraffin-embedded sections were stained with antibodies against AQP3 (Millipore), human EpCAM (eFluor 660; eBioscience), turbo-GFP (Origene), or F4/80 (allophycocyanin [APC]; Biolegend).

Techniques: Transfection, Multiple Displacement Amplification, shRNA

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: AQP3 knockdown abrogates spontaneous metastasis to the lungs. (A to F) Control or AQP3 KD MDA-MB-231 cells (turbo-GFP-expressing control or AQP3 shRNA; 1.0 × 10 6 cells) were orthotopically injected into SCID mice. Primary tumors and lungs were

Article Snippet: For AQP3 staining and visualization of macrophage infiltration, frozen or paraffin-embedded sections were stained with antibodies against AQP3 (Millipore), human EpCAM (eFluor 660; eBioscience), turbo-GFP (Origene), or F4/80 (allophycocyanin [APC]; Biolegend).

Techniques: Multiple Displacement Amplification, Expressing, shRNA, Injection, Mouse Assay

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: AQP3 overexpression increases H 2 O 2 uptake and cell migration upon CXCL12 stimulation. The error bars indicate SE. (A to F) MDA-MB-231 cells were transfected with the vector pCMV6 (empty [Emp]) or human AQP3-expressing pCMV6 (AQP3; 1 to 10 ng for 1.5 ×

Article Snippet: For AQP3 staining and visualization of macrophage infiltration, frozen or paraffin-embedded sections were stained with antibodies against AQP3 (Millipore), human EpCAM (eFluor 660; eBioscience), turbo-GFP (Origene), or F4/80 (allophycocyanin [APC]; Biolegend).

Techniques: Over Expression, Migration, Multiple Displacement Amplification, Transfection, Plasmid Preparation, Expressing

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: CXCL12-induced directional migration requires AQP3 and H 2 O 2 uptake. (A to F) MDA-MB-231 and DU4475 cells were transfected with control or AQP3 siRNA. The error bars indicate SE. (A) The chemotaxis efficiency of control or AQP3 KD MDA-MB-231 and DU4475

Article Snippet: The supernatant (10,000 rpm; 10 min; 4°C) was used for immunoblotting with antibodies against AQP3 (Abcam), Na+ /K+-ATPase (Millipore), CD98 (Abcam), or Nox2 (Millipore).

Techniques: Migration, Multiple Displacement Amplification, Transfection, Chemotaxis Assay

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: AQP3-dependent PTEN/PTP1B oxidation and Akt phosphorylation. (A to E) MDA-MB-231 cells were infected with lentiviral control or AQP3 shRNA, and DU4475 cells were transfected with control or AQP3 siRNA. (A) Immunoblot analysis of PTEN oxidation. Control

Article Snippet: The supernatant (10,000 rpm; 10 min; 4°C) was used for immunoblotting with antibodies against AQP3 (Abcam), Na+ /K+-ATPase (Millipore), CD98 (Abcam), or Nox2 (Millipore).

Techniques: Multiple Displacement Amplification, Infection, shRNA, Transfection

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: Involvement of AQP3 expression in breast cancer cell migration in vivo . The error bars indicate SE. (A to D) MDA-MB-231 cells (8.0 × 10 5 cells) were intravenously injected into SCID mice, and the numbers of cells in the lungs after 24 h were analyzed

Article Snippet: The supernatant (10,000 rpm; 10 min; 4°C) was used for immunoblotting with antibodies against AQP3 (Abcam), Na+ /K+-ATPase (Millipore), CD98 (Abcam), or Nox2 (Millipore).

Techniques: Expressing, Migration, In Vivo, Multiple Displacement Amplification, Injection, Mouse Assay

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: CXCL12-induced H 2 O 2 regulates PTP1B/PTEN oxidation and Akt activation. (A to C, E, and F) MDA-MB-231 cells were infected with lentiviral control or AQP3 shRNA. (A and B) The effects of DPI (20 μM; 30 min) or catalase (2,000 U/ml) on PTEN (A) or

Article Snippet: The supernatant (10,000 rpm; 10 min; 4°C) was used for immunoblotting with antibodies against AQP3 (Abcam), Na+ /K+-ATPase (Millipore), CD98 (Abcam), or Nox2 (Millipore).

Techniques: Activation Assay, Multiple Displacement Amplification, Infection, shRNA

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: Transport of CXCL12-induced H 2 O 2 into breast cancer cells through AQP3. (A) The mRNA levels of AQP3 in control (si-control)- or AQP3-siRNA (si-AQP3)-transfected MDA-MB-231 (top) and DU4475 (bottom) cells and in lentiviral control (sh-control)- or AQP3-shRNA

Article Snippet: The supernatant (10,000 rpm; 10 min; 4°C) was used for immunoblotting with antibodies against AQP3 (Abcam), Na+ /K+-ATPase (Millipore), CD98 (Abcam), or Nox2 (Millipore).

Techniques: Transfection, Multiple Displacement Amplification, shRNA

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: AQP3 knockdown abrogates spontaneous metastasis to the lungs. (A to F) Control or AQP3 KD MDA-MB-231 cells (turbo-GFP-expressing control or AQP3 shRNA; 1.0 × 10 6 cells) were orthotopically injected into SCID mice. Primary tumors and lungs were

Article Snippet: The supernatant (10,000 rpm; 10 min; 4°C) was used for immunoblotting with antibodies against AQP3 (Abcam), Na+ /K+-ATPase (Millipore), CD98 (Abcam), or Nox2 (Millipore).

Techniques: Multiple Displacement Amplification, Expressing, shRNA, Injection, Mouse Assay

Journal: Molecular and Cellular Biology

Article Title: Aquaporin-3 Controls Breast Cancer Cell Migration by Regulating Hydrogen Peroxide Transport and Its Downstream Cell Signaling

doi: 10.1128/MCB.00971-15

Figure Lengend Snippet: AQP3 overexpression increases H 2 O 2 uptake and cell migration upon CXCL12 stimulation. The error bars indicate SE. (A to F) MDA-MB-231 cells were transfected with the vector pCMV6 (empty [Emp]) or human AQP3-expressing pCMV6 (AQP3; 1 to 10 ng for 1.5 ×

Article Snippet: The supernatant (10,000 rpm; 10 min; 4°C) was used for immunoblotting with antibodies against AQP3 (Abcam), Na+ /K+-ATPase (Millipore), CD98 (Abcam), or Nox2 (Millipore).

Techniques: Over Expression, Migration, Multiple Displacement Amplification, Transfection, Plasmid Preparation, Expressing

Journal: Cell Death Discovery

Article Title: Aquaporin 3 facilitates chemoresistance in gastric cancer cells to cisplatin via autophagy

doi: 10.1038/cddiscovery.2016.87

Figure Lengend Snippet: Aquaporin 3 (AQP3) enhances autophagy in gastric cancer cells. ( a ) AQP3 overexpression induced the conversion of LC3-I to LC3-II in AGS cells, and AQP3 knockdown inhibited this conversion in MGC803 and SGC7901 cells ( b , * P

Article Snippet: Western blotting Western blotting was performed to analyze the expression of proteins according to our published methods., We used antibodies against AQP3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), LC3 A/B (Cell Signaling Technology) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Beyotime Institute of Biotechnology).

Techniques: Over Expression

Journal: International Journal of Molecular Sciences

Article Title: AQP3 Increases Intercellular Cohesion in NSCLC A549 Cell Spheroids through Exploratory Cell Protrusions

doi: 10.3390/ijms22084287

Figure Lengend Snippet: A549 cell aggregation is an active process. ( A ) The effect of ROCK on A549 cell aggregation. The phase-contrast micrographs show the morphologies of the A549 cells following treatment with a ROCK inhibitor, Y-27632, and the Rho activator Ⅱ. ( B ) The effects of AQP3 knockdown with siRNA on actomyosin cytoskeleton remodeling. A549 cells were stained with anti-AQP3 antibody, followed by Fluorescein-conjugated antibody (green). The actin microfilaments were stained with rhodamine-conjugated phalloidin (red), and the nuclei were stained with DAPI (blue). ( C ) The effects of AQP3 knockdown with siRNA on the apoptosis signaling pathway. Twenty-four h following siRNA transfection in 2D culture condition, the cells were further incubated in the 2D or 3D culture condition. Then, cells were harvested to confirm the effect of AQP3 knockdown on the apoptotic molecular signatures using Western blotting. α-tubulin was used as an internal control.

Article Snippet: In short, the cells on coverslips were fixed with 4% paraformaldehyde for 10 min and permeabilized with 0.15% Triton-X 100 for 5 min. Then, the cells were blocked for 1 h with the blocking solution of 3% bovine serum albumin in PBS and incubated with the primary antibody against AQP3 for 2 h at room temperature.

Techniques: Staining, Transfection, Incubation, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: AQP3 Increases Intercellular Cohesion in NSCLC A549 Cell Spheroids through Exploratory Cell Protrusions

doi: 10.3390/ijms22084287

Figure Lengend Snippet: The effect of AQP3 on spatiotemporal dynamics of protrusions. ( A ) Polymerase chain reaction and ( B ) Quantitative real-time reverse transcription-polymerase chain reaction of the transcript levels of the organic hydroxyl transport genes. The data shown here represent three independent experiments, and the values represent the mean ± SEM of triplicate samples. The level of each mRNA was normalized to that of the GAPDH mRNA in the same sample and presented as the fold-change over that of the 2D culture control cells. The differences in expression levels were evaluated for significance using two-tailed t-tests with unequal variance. * p

Article Snippet: In short, the cells on coverslips were fixed with 4% paraformaldehyde for 10 min and permeabilized with 0.15% Triton-X 100 for 5 min. Then, the cells were blocked for 1 h with the blocking solution of 3% bovine serum albumin in PBS and incubated with the primary antibody against AQP3 for 2 h at room temperature.

Techniques: Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing, Two Tailed Test