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Santa Cruz Biotechnology v atpase
Quantitative analyses of the expression levels of <t>CD147</t> and <t>V-ATPase</t> in breast cancer samples. Comparison of the expression levels of (A) CD147 and (B) V-ATPase between the chemotherapy-sensitive and -resistant groups. (C) Correlation between the H-scores of CD147 and V-ATPase in breast invasive ductal cancer. ***P
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1) Product Images from "Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity"

Article Title: Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity

Journal: Oncology Letters

doi: 10.3892/ol.2018.8199

Quantitative analyses of the expression levels of CD147 and V-ATPase in breast cancer samples. Comparison of the expression levels of (A) CD147 and (B) V-ATPase between the chemotherapy-sensitive and -resistant groups. (C) Correlation between the H-scores of CD147 and V-ATPase in breast invasive ductal cancer. ***P
Figure Legend Snippet: Quantitative analyses of the expression levels of CD147 and V-ATPase in breast cancer samples. Comparison of the expression levels of (A) CD147 and (B) V-ATPase between the chemotherapy-sensitive and -resistant groups. (C) Correlation between the H-scores of CD147 and V-ATPase in breast invasive ductal cancer. ***P

Techniques Used: Expressing

Immunohistochemical observation of CD147 and V-ATPase expression in chemotherapy-sensitive or -resistant breast invasive ductal cancer patients (magnification, ×400). Chemotherapy-sensitive patients exhibited moderate expression of (A) CD147 and (B) V-ATPase in the cell membrane. Chemotherapy-resistant patients exhibited high expression of (C) CD147 in the cell membrane and (D) V-ATPase mainly in the cell membrane and cytoplasm. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase.
Figure Legend Snippet: Immunohistochemical observation of CD147 and V-ATPase expression in chemotherapy-sensitive or -resistant breast invasive ductal cancer patients (magnification, ×400). Chemotherapy-sensitive patients exhibited moderate expression of (A) CD147 and (B) V-ATPase in the cell membrane. Chemotherapy-resistant patients exhibited high expression of (C) CD147 in the cell membrane and (D) V-ATPase mainly in the cell membrane and cytoplasm. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase.

Techniques Used: Immunohistochemistry, Expressing

Immunofluorescence was used to detect CD147 and V-ATPase expression and subcellular localization in the four transfected cell lines. Experiments were repeated at least three times, and a representative experiment is shown. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase; CON, control; CD, CD147 overexpression; si, CD147 knockdown. Blue represents nuclear staining; red represents CD147 staining; yellow represents V-ATPase staining.
Figure Legend Snippet: Immunofluorescence was used to detect CD147 and V-ATPase expression and subcellular localization in the four transfected cell lines. Experiments were repeated at least three times, and a representative experiment is shown. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase; CON, control; CD, CD147 overexpression; si, CD147 knockdown. Blue represents nuclear staining; red represents CD147 staining; yellow represents V-ATPase staining.

Techniques Used: Immunofluorescence, Expressing, Transfection, Over Expression, Staining

2) Product Images from "Whole Transcriptome Analysis of Renal Intercalated Cells Predicts Lipopolysaccharide Mediated Inhibition of Retinoid X Receptor alpha Function"

Article Title: Whole Transcriptome Analysis of Renal Intercalated Cells Predicts Lipopolysaccharide Mediated Inhibition of Retinoid X Receptor alpha Function

Journal: Scientific Reports

doi: 10.1038/s41598-018-36921-z

( A ) Triple immunofluorescence labeling of a kidney section from AQP2-cre + tdTomato + (PC reporter) mice showing: AQP2 (red cells, upper left);. V-ATPase E1 (green cells, ICs, upper right); tdTomato (blue cells, lower left) staining in inner medulla. On the merged image (lower right) most AQP2 cells were also positive for tdTomato (purple cells, white arrows) while some were negative for tdTomato (red cells, white arrowheads). The dashed box contains a tubule imaged from another kidney section that contained an isolated cell that was positive for both AQP2 and V-ATPAse E1 (Yellow cell, asterisk). Magnification 40X, confocal microscopy. ( B ) Triple immunofluorescence labeling of a kidney section from V-ATPase B1-cre + tdTomato + (IC reporter) mice showing: V-ATPase B1 (red cells, upper left). tdTomato (green cells, upper right) and nuclei (blue, lower left) staining in inner medulla. On the merged image (lower right) most V-ATPase B1 positive cells were also tdTomato positive (yellow cells, white arrows) while a few cells were tdTomato positive but were V-ATPase B1 negative (green cells, white arrowheads). Magnification 20X, immunofluorescent microscopy.
Figure Legend Snippet: ( A ) Triple immunofluorescence labeling of a kidney section from AQP2-cre + tdTomato + (PC reporter) mice showing: AQP2 (red cells, upper left);. V-ATPase E1 (green cells, ICs, upper right); tdTomato (blue cells, lower left) staining in inner medulla. On the merged image (lower right) most AQP2 cells were also positive for tdTomato (purple cells, white arrows) while some were negative for tdTomato (red cells, white arrowheads). The dashed box contains a tubule imaged from another kidney section that contained an isolated cell that was positive for both AQP2 and V-ATPAse E1 (Yellow cell, asterisk). Magnification 40X, confocal microscopy. ( B ) Triple immunofluorescence labeling of a kidney section from V-ATPase B1-cre + tdTomato + (IC reporter) mice showing: V-ATPase B1 (red cells, upper left). tdTomato (green cells, upper right) and nuclei (blue, lower left) staining in inner medulla. On the merged image (lower right) most V-ATPase B1 positive cells were also tdTomato positive (yellow cells, white arrows) while a few cells were tdTomato positive but were V-ATPase B1 negative (green cells, white arrowheads). Magnification 20X, immunofluorescent microscopy.

Techniques Used: Immunofluorescence, Labeling, Mouse Assay, Staining, Isolation, Confocal Microscopy, Microscopy

3) Product Images from "Whole Transcriptome Analysis of Renal Intercalated Cells Predicts Lipopolysaccharide Mediated Inhibition of Retinoid X Receptor alpha Function"

Article Title: Whole Transcriptome Analysis of Renal Intercalated Cells Predicts Lipopolysaccharide Mediated Inhibition of Retinoid X Receptor alpha Function

Journal: Scientific Reports

doi: 10.1038/s41598-018-36921-z

( A ) Triple immunofluorescence labeling of a kidney section from AQP2-cre + tdTomato + (PC reporter) mice showing: AQP2 (red cells, upper left);. V-ATPase E1 (green cells, ICs, upper right); tdTomato (blue cells, lower left) staining in inner medulla. On the merged image (lower right) most AQP2 cells were also positive for tdTomato (purple cells, white arrows) while some were negative for tdTomato (red cells, white arrowheads). The dashed box contains a tubule imaged from another kidney section that contained an isolated cell that was positive for both AQP2 and V-ATPAse E1 (Yellow cell, asterisk). Magnification 40X, confocal microscopy. ( B ) Triple immunofluorescence labeling of a kidney section from V-ATPase B1-cre + tdTomato + (IC reporter) mice showing: V-ATPase B1 (red cells, upper left). tdTomato (green cells, upper right) and nuclei (blue, lower left) staining in inner medulla. On the merged image (lower right) most V-ATPase B1 positive cells were also tdTomato positive (yellow cells, white arrows) while a few cells were tdTomato positive but were V-ATPase B1 negative (green cells, white arrowheads). Magnification 20X, immunofluorescent microscopy.
Figure Legend Snippet: ( A ) Triple immunofluorescence labeling of a kidney section from AQP2-cre + tdTomato + (PC reporter) mice showing: AQP2 (red cells, upper left);. V-ATPase E1 (green cells, ICs, upper right); tdTomato (blue cells, lower left) staining in inner medulla. On the merged image (lower right) most AQP2 cells were also positive for tdTomato (purple cells, white arrows) while some were negative for tdTomato (red cells, white arrowheads). The dashed box contains a tubule imaged from another kidney section that contained an isolated cell that was positive for both AQP2 and V-ATPAse E1 (Yellow cell, asterisk). Magnification 40X, confocal microscopy. ( B ) Triple immunofluorescence labeling of a kidney section from V-ATPase B1-cre + tdTomato + (IC reporter) mice showing: V-ATPase B1 (red cells, upper left). tdTomato (green cells, upper right) and nuclei (blue, lower left) staining in inner medulla. On the merged image (lower right) most V-ATPase B1 positive cells were also tdTomato positive (yellow cells, white arrows) while a few cells were tdTomato positive but were V-ATPase B1 negative (green cells, white arrowheads). Magnification 20X, immunofluorescent microscopy.

Techniques Used: Immunofluorescence, Labeling, Mouse Assay, Staining, Isolation, Confocal Microscopy, Microscopy

4) Product Images from "Nicotine Affects Bone Resorption and Suppresses the Expression of Cathepsin K, MMP-9 and Vacuolar-Type H+-ATPase d2 and Actin Organization in Osteoclasts"

Article Title: Nicotine Affects Bone Resorption and Suppresses the Expression of Cathepsin K, MMP-9 and Vacuolar-Type H+-ATPase d2 and Actin Organization in Osteoclasts

Journal: PLoS ONE

doi: 10.1371/journal.pone.0059402

Effect of nicotine on protein expression of CA II, cathepsin K, MMP-9, and V-ATPase d2. RAW264.7 cells were cultured in differentiation medium with 0 (control), 10 −5 , 10 −4 , or 10 −3 M nicotine for 5 days. The protein expression of CA II, cathepsin K, MMP-9, and V-ATPase was determined by Western blotting.
Figure Legend Snippet: Effect of nicotine on protein expression of CA II, cathepsin K, MMP-9, and V-ATPase d2. RAW264.7 cells were cultured in differentiation medium with 0 (control), 10 −5 , 10 −4 , or 10 −3 M nicotine for 5 days. The protein expression of CA II, cathepsin K, MMP-9, and V-ATPase was determined by Western blotting.

Techniques Used: Expressing, Cell Culture, Western Blot

Effect of btx and/or nicotine on the expression of CA II, cathepsin K, and MMP-9 and V-ATPase d2. RAW264.7 cells were cultured in differentiation medium without drug (control), 50 µM btx, 10 −3 M nicotine, or 50 µM btx and 10 −3 M nicotine for 5 days. The mRNA expression of CA II (a), cathepsin K (b), MMP-9 (c), and V-ATPase d2 (d) was determined by real-time PCR. Cells were cultured in differentiation medium without drug (control), 50 µM btx, 10 −3 M nicotine, or 50 µM btx and 10 −3 M nicotine for 5 days. The protein expression of CA II, cathepsin K, MMP-9, and V-ATPase d2 was determined by Western blotting (e). Data are shown as the mean±S.D., n = 3 independent experiments. *p
Figure Legend Snippet: Effect of btx and/or nicotine on the expression of CA II, cathepsin K, and MMP-9 and V-ATPase d2. RAW264.7 cells were cultured in differentiation medium without drug (control), 50 µM btx, 10 −3 M nicotine, or 50 µM btx and 10 −3 M nicotine for 5 days. The mRNA expression of CA II (a), cathepsin K (b), MMP-9 (c), and V-ATPase d2 (d) was determined by real-time PCR. Cells were cultured in differentiation medium without drug (control), 50 µM btx, 10 −3 M nicotine, or 50 µM btx and 10 −3 M nicotine for 5 days. The protein expression of CA II, cathepsin K, MMP-9, and V-ATPase d2 was determined by Western blotting (e). Data are shown as the mean±S.D., n = 3 independent experiments. *p

Techniques Used: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot

Effect of btx and/or nicotine on the expression of CA II, cathepsin K, and MMP-9 and V-ATPase d2. Bone marrow cells were cultured in medium containing macrophage colony-stimulating factor (50 ng/ml), and RANKL (50 ng/ml) without drug (control), 50 µM btx, 10 −3 M nicotine, or 50 µM btx, and 10 −3 M nicotine for 5 days. The mRNA expression of CA II (a), cathepsin K (b), MMP-9 (c), and V-ATPase d2 (d) was determined by real-time PCR. The protein expression of CA II, cathepsin K, MMP-9, and V-ATPase d2 was determined by Western blotting (e).Data are shown as the mean±S.D., n = 3 independent experiments. *p
Figure Legend Snippet: Effect of btx and/or nicotine on the expression of CA II, cathepsin K, and MMP-9 and V-ATPase d2. Bone marrow cells were cultured in medium containing macrophage colony-stimulating factor (50 ng/ml), and RANKL (50 ng/ml) without drug (control), 50 µM btx, 10 −3 M nicotine, or 50 µM btx, and 10 −3 M nicotine for 5 days. The mRNA expression of CA II (a), cathepsin K (b), MMP-9 (c), and V-ATPase d2 (d) was determined by real-time PCR. The protein expression of CA II, cathepsin K, MMP-9, and V-ATPase d2 was determined by Western blotting (e).Data are shown as the mean±S.D., n = 3 independent experiments. *p

Techniques Used: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot

Effect of nicotine on mRNA expression of CA II, cathepsin K, MMP-9, and V-ATPase d2. RAW264.7 cells were cultured in differentiation medium with 0 (control), 10 −5 , 10 −4 , or 10 −3 M nicotine for up to 7 days. The mRNA expression of CA II (a), cathepsin K (b), MMP-9 (c), and V-ATPase d2 (d) was determined using real-time PCR on days 1, 3, 5, and 7 of culture. Data are shown as the mean±S.D., n = 3 independent experiments. *p
Figure Legend Snippet: Effect of nicotine on mRNA expression of CA II, cathepsin K, MMP-9, and V-ATPase d2. RAW264.7 cells were cultured in differentiation medium with 0 (control), 10 −5 , 10 −4 , or 10 −3 M nicotine for up to 7 days. The mRNA expression of CA II (a), cathepsin K (b), MMP-9 (c), and V-ATPase d2 (d) was determined using real-time PCR on days 1, 3, 5, and 7 of culture. Data are shown as the mean±S.D., n = 3 independent experiments. *p

Techniques Used: Expressing, Cell Culture, Real-time Polymerase Chain Reaction

5) Product Images from "Low Na, High K Diet and the Role of Aldosterone in BK-Mediated K Excretion"

Article Title: Low Na, High K Diet and the Role of Aldosterone in BK-Mediated K Excretion

Journal: PLoS ONE

doi: 10.1371/journal.pone.0115515

The effect of aldo on BK-α expression in the distal nephron of LNaHK mice. Representative images (A) and bar plots summarizing intensity of fluorescent immunohistochemical staining of apical (B) and total (C) BK-α in cortical kidney sections of sham, ADX-LA, and ADX-HA mice. BK-α (green) was co-stained with V-ATPase (red), a marker of intercalated cells in the connecting tubule and collecting duct. Merged images also contain Hoechst nuclear stain (blue). * P
Figure Legend Snippet: The effect of aldo on BK-α expression in the distal nephron of LNaHK mice. Representative images (A) and bar plots summarizing intensity of fluorescent immunohistochemical staining of apical (B) and total (C) BK-α in cortical kidney sections of sham, ADX-LA, and ADX-HA mice. BK-α (green) was co-stained with V-ATPase (red), a marker of intercalated cells in the connecting tubule and collecting duct. Merged images also contain Hoechst nuclear stain (blue). * P

Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Staining, Marker

6) Product Images from "Mutation of Prkar1a Causes Osteoblast Neoplasia Driven by Dysregulation of Protein Kinase A"

Article Title: Mutation of Prkar1a Causes Osteoblast Neoplasia Driven by Dysregulation of Protein Kinase A

Journal: Molecular Endocrinology

doi: 10.1210/me.2007-0369

Gross and Immunohistochemical Characterization of Bone Tumors in Prkar1a +/− Mice A, X-ray of a mouse with early tail tumors showing effacement of the normal vertebral bone; B, tail of WT ( left ) and Prkar1a +/− ( right ) mice stained for bone and cartilage with Alizarin red (bone) and Alcian blue (cartilage); C–E, immunohistochemical analysis of bone tumors for Runx2 (C), a marker of early osteoblast differentiation; osteocalcin (D), a marker of late osteoblast maturation; and V-ATPase (E), a marker for osteoclasts ( black arrows ); F, quantitation of osteoclast numbers per HPF in the bone tumors. The graph shows the average counts from 15 HPF from two WT and two tumor tails. In C and D, tumor cells are shown with black arrows , whereas normal staining osteoblasts are shown with black arrowheads .
Figure Legend Snippet: Gross and Immunohistochemical Characterization of Bone Tumors in Prkar1a +/− Mice A, X-ray of a mouse with early tail tumors showing effacement of the normal vertebral bone; B, tail of WT ( left ) and Prkar1a +/− ( right ) mice stained for bone and cartilage with Alizarin red (bone) and Alcian blue (cartilage); C–E, immunohistochemical analysis of bone tumors for Runx2 (C), a marker of early osteoblast differentiation; osteocalcin (D), a marker of late osteoblast maturation; and V-ATPase (E), a marker for osteoclasts ( black arrows ); F, quantitation of osteoclast numbers per HPF in the bone tumors. The graph shows the average counts from 15 HPF from two WT and two tumor tails. In C and D, tumor cells are shown with black arrows , whereas normal staining osteoblasts are shown with black arrowheads .

Techniques Used: Immunohistochemistry, Mouse Assay, Staining, Marker, Quantitation Assay

7) Product Images from "Mutation of Prkar1a Causes Osteoblast Neoplasia Driven by Dysregulation of Protein Kinase A"

Article Title: Mutation of Prkar1a Causes Osteoblast Neoplasia Driven by Dysregulation of Protein Kinase A

Journal: Molecular Endocrinology

doi: 10.1210/me.2007-0369

Gross and Immunohistochemical Characterization of Bone Tumors in Prkar1a +/− Mice A, X-ray of a mouse with early tail tumors showing effacement of the normal vertebral bone; B, tail of WT ( left ) and Prkar1a +/− ( right ) mice stained for bone and cartilage with Alizarin red (bone) and Alcian blue (cartilage); C–E, immunohistochemical analysis of bone tumors for Runx2 (C), a marker of early osteoblast differentiation; osteocalcin (D), a marker of late osteoblast maturation; and V-ATPase (E), a marker for osteoclasts ( black arrows ); F, quantitation of osteoclast numbers per HPF in the bone tumors. The graph shows the average counts from 15 HPF from two WT and two tumor tails. In C and D, tumor cells are shown with black arrows , whereas normal staining osteoblasts are shown with black arrowheads .
Figure Legend Snippet: Gross and Immunohistochemical Characterization of Bone Tumors in Prkar1a +/− Mice A, X-ray of a mouse with early tail tumors showing effacement of the normal vertebral bone; B, tail of WT ( left ) and Prkar1a +/− ( right ) mice stained for bone and cartilage with Alizarin red (bone) and Alcian blue (cartilage); C–E, immunohistochemical analysis of bone tumors for Runx2 (C), a marker of early osteoblast differentiation; osteocalcin (D), a marker of late osteoblast maturation; and V-ATPase (E), a marker for osteoclasts ( black arrows ); F, quantitation of osteoclast numbers per HPF in the bone tumors. The graph shows the average counts from 15 HPF from two WT and two tumor tails. In C and D, tumor cells are shown with black arrows , whereas normal staining osteoblasts are shown with black arrowheads .

Techniques Used: Immunohistochemistry, Mouse Assay, Staining, Marker, Quantitation Assay

8) Product Images from "Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity"

Article Title: Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity

Journal: Oncology Letters

doi: 10.3892/ol.2018.8199

Quantitative analyses of the expression levels of CD147 and V-ATPase in breast cancer samples. Comparison of the expression levels of (A) CD147 and (B) V-ATPase between the chemotherapy-sensitive and -resistant groups. (C) Correlation between the H-scores of CD147 and V-ATPase in breast invasive ductal cancer. ***P
Figure Legend Snippet: Quantitative analyses of the expression levels of CD147 and V-ATPase in breast cancer samples. Comparison of the expression levels of (A) CD147 and (B) V-ATPase between the chemotherapy-sensitive and -resistant groups. (C) Correlation between the H-scores of CD147 and V-ATPase in breast invasive ductal cancer. ***P

Techniques Used: Expressing

Immunohistochemical observation of CD147 and V-ATPase expression in chemotherapy-sensitive or -resistant breast invasive ductal cancer patients (magnification, ×400). Chemotherapy-sensitive patients exhibited moderate expression of (A) CD147 and (B) V-ATPase in the cell membrane. Chemotherapy-resistant patients exhibited high expression of (C) CD147 in the cell membrane and (D) V-ATPase mainly in the cell membrane and cytoplasm. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase.
Figure Legend Snippet: Immunohistochemical observation of CD147 and V-ATPase expression in chemotherapy-sensitive or -resistant breast invasive ductal cancer patients (magnification, ×400). Chemotherapy-sensitive patients exhibited moderate expression of (A) CD147 and (B) V-ATPase in the cell membrane. Chemotherapy-resistant patients exhibited high expression of (C) CD147 in the cell membrane and (D) V-ATPase mainly in the cell membrane and cytoplasm. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase.

Techniques Used: Immunohistochemistry, Expressing

Immunofluorescence was used to detect CD147 and V-ATPase expression and subcellular localization in the four transfected cell lines. Experiments were repeated at least three times, and a representative experiment is shown. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase; CON, control; CD, CD147 overexpression; si, CD147 knockdown. Blue represents nuclear staining; red represents CD147 staining; yellow represents V-ATPase staining.
Figure Legend Snippet: Immunofluorescence was used to detect CD147 and V-ATPase expression and subcellular localization in the four transfected cell lines. Experiments were repeated at least three times, and a representative experiment is shown. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase; CON, control; CD, CD147 overexpression; si, CD147 knockdown. Blue represents nuclear staining; red represents CD147 staining; yellow represents V-ATPase staining.

Techniques Used: Immunofluorescence, Expressing, Transfection, Over Expression, Staining

Related Articles

Incubation:

Article Title: Whole Transcriptome Analysis of Renal Intercalated Cells Predicts Lipopolysaccharide Mediated Inhibition of Retinoid X Receptor alpha Function
Article Snippet: .. V-ATPase-cre mice : In this mice, to visualize, V-ATPase, goat anti-mouse V-ATPase B1/B2 antibody (E20) (sc21209, 1:50 dilution, Santa Cruz, CA), to visualize tdTomato, anti-RFP-FITC conjugated antibody (ab34764, 1:200 dilution, abcam, MA) were incubated overnight at 4 °C. ..

Article Title: Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity
Article Snippet: .. Subsequent to heating for 20 min at 100°C in a microwave oven for antigen retrieval and blocking with normal rabbit serum for 20 min at room temperature (cat. no. ab166640; Abcam), the sections were incubated with primary antibodies at 4°C overnight, including CD147 (1:200; cat. no. ab212856; Abcam) and V-ATPase (1:200; cat. no. sc-69088; Santa Cruz Biotechnology, Inc. Dallas, TX, USA), or with PBS as the negative control. .. A 2-step Plus Poly-HRP Anti-Mouse/Goat IgG detection system (OriGene Technologies, Inc., Beijing, China) was then applied according to the manufacturer's protocol, followed by DAB visualization.

Article Title: Nicotine Affects Bone Resorption and Suppresses the Expression of Cathepsin K, MMP-9 and Vacuolar-Type H+-ATPase d2 and Actin Organization in Osteoclasts
Article Snippet: .. Gels were run at 150 V for 60 min. Gel-separated proteins were transferred to a membrane using a semidry electrotransfer unit with a continuous buffer system consisting 39 mM glycine, 48 mM Tris, 0.0375% SDS, and 20% (v/v) methanol at a constant amperage of 0.8 mA/cm2 for 60–90 min. On completion of the transfer, the transfer membrane was treated with 25% (v/v) blocking reagent in Tris-buffered saline (TBS) (10 mM Tris, 145 mM NaCl, pH 7.4) at 4°C for 18 h. The sheet was washed in TBS containing Tween 20 (TBS-Tween) and then incubated at room temperature for 90 min with rabbit or goat biotin-labeled polyclonal IgG antibodies against CA II, MMP-9, cathepsin K, V-ATPase, and β-actin (all from Santa Cruz Biotechnology, Santa Cruz, CA, USA). .. Primary antibodies were diluted 1∶200 in distilled water containing 10% (v/v) blocking reagent. β-actin was used as an internal standard.

Article Title: Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity
Article Snippet: .. Subsequent to washing with PBS, the cells were incubated for 2 h at room temperature with primary antibodies against CD147 (1:200; cat. no. ab212856; Abcam) and V-ATPase (1:100; cat. no. sc-69088; Santa Cruz Biotechnology, Inc. Dallas, TX, USA), followed by incubation with secondary antibodies, including anti-mouse Cy3 (1:200; cat. no. AP192C; EMD Millipore, Billerica, MA, USA) or anti-goat FITC (1:500; cat. no. ab6881; Abcam) antibodies for 1 h at room temperature. .. Cells were then rinsed with PBS for three times and mounted in a ProLong Gold Antifade reagent with DAPI (Invitrogen; Thermo Fisher Scientific, Inc.).

Negative Control:

Article Title: Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity
Article Snippet: .. Subsequent to heating for 20 min at 100°C in a microwave oven for antigen retrieval and blocking with normal rabbit serum for 20 min at room temperature (cat. no. ab166640; Abcam), the sections were incubated with primary antibodies at 4°C overnight, including CD147 (1:200; cat. no. ab212856; Abcam) and V-ATPase (1:200; cat. no. sc-69088; Santa Cruz Biotechnology, Inc. Dallas, TX, USA), or with PBS as the negative control. .. A 2-step Plus Poly-HRP Anti-Mouse/Goat IgG detection system (OriGene Technologies, Inc., Beijing, China) was then applied according to the manufacturer's protocol, followed by DAB visualization.

Electrotransfer:

Article Title: Nicotine Affects Bone Resorption and Suppresses the Expression of Cathepsin K, MMP-9 and Vacuolar-Type H+-ATPase d2 and Actin Organization in Osteoclasts
Article Snippet: .. Gels were run at 150 V for 60 min. Gel-separated proteins were transferred to a membrane using a semidry electrotransfer unit with a continuous buffer system consisting 39 mM glycine, 48 mM Tris, 0.0375% SDS, and 20% (v/v) methanol at a constant amperage of 0.8 mA/cm2 for 60–90 min. On completion of the transfer, the transfer membrane was treated with 25% (v/v) blocking reagent in Tris-buffered saline (TBS) (10 mM Tris, 145 mM NaCl, pH 7.4) at 4°C for 18 h. The sheet was washed in TBS containing Tween 20 (TBS-Tween) and then incubated at room temperature for 90 min with rabbit or goat biotin-labeled polyclonal IgG antibodies against CA II, MMP-9, cathepsin K, V-ATPase, and β-actin (all from Santa Cruz Biotechnology, Santa Cruz, CA, USA). .. Primary antibodies were diluted 1∶200 in distilled water containing 10% (v/v) blocking reagent. β-actin was used as an internal standard.

Blocking Assay:

Article Title: Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity
Article Snippet: .. Subsequent to heating for 20 min at 100°C in a microwave oven for antigen retrieval and blocking with normal rabbit serum for 20 min at room temperature (cat. no. ab166640; Abcam), the sections were incubated with primary antibodies at 4°C overnight, including CD147 (1:200; cat. no. ab212856; Abcam) and V-ATPase (1:200; cat. no. sc-69088; Santa Cruz Biotechnology, Inc. Dallas, TX, USA), or with PBS as the negative control. .. A 2-step Plus Poly-HRP Anti-Mouse/Goat IgG detection system (OriGene Technologies, Inc., Beijing, China) was then applied according to the manufacturer's protocol, followed by DAB visualization.

Article Title: Nicotine Affects Bone Resorption and Suppresses the Expression of Cathepsin K, MMP-9 and Vacuolar-Type H+-ATPase d2 and Actin Organization in Osteoclasts
Article Snippet: .. Gels were run at 150 V for 60 min. Gel-separated proteins were transferred to a membrane using a semidry electrotransfer unit with a continuous buffer system consisting 39 mM glycine, 48 mM Tris, 0.0375% SDS, and 20% (v/v) methanol at a constant amperage of 0.8 mA/cm2 for 60–90 min. On completion of the transfer, the transfer membrane was treated with 25% (v/v) blocking reagent in Tris-buffered saline (TBS) (10 mM Tris, 145 mM NaCl, pH 7.4) at 4°C for 18 h. The sheet was washed in TBS containing Tween 20 (TBS-Tween) and then incubated at room temperature for 90 min with rabbit or goat biotin-labeled polyclonal IgG antibodies against CA II, MMP-9, cathepsin K, V-ATPase, and β-actin (all from Santa Cruz Biotechnology, Santa Cruz, CA, USA). .. Primary antibodies were diluted 1∶200 in distilled water containing 10% (v/v) blocking reagent. β-actin was used as an internal standard.

Mouse Assay:

Article Title: Whole Transcriptome Analysis of Renal Intercalated Cells Predicts Lipopolysaccharide Mediated Inhibition of Retinoid X Receptor alpha Function
Article Snippet: .. V-ATPase-cre mice : In this mice, to visualize, V-ATPase, goat anti-mouse V-ATPase B1/B2 antibody (E20) (sc21209, 1:50 dilution, Santa Cruz, CA), to visualize tdTomato, anti-RFP-FITC conjugated antibody (ab34764, 1:200 dilution, abcam, MA) were incubated overnight at 4 °C. ..

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    Santa Cruz Biotechnology antibodies against v atpase g1
    Bafilomycin A1 repress stemness features of GBM neurospheres A. Nestin expression was evaluated by confocal immunofluorescence in GBM neurospheres incubated for 48 hours with vehicle or different concentrations of bafilomycin A1 and quantified in multiple z-stacks by ROI. **, p = 0.007; ***, p = 0.0006 by Mann-Whitney U test. Bars, mean ±SEM ( n = 5). Left , Representative maximum intensity projection images of neurospheres stained for the indicated antibody. B. , C. Gene expression evaluation of ATP6V1G1, CD133 and NESTIN, or of transcription factors involved in neurodevelopment and reprogramming (SOX2, OLIG2, SALL2 and POU3F2) in GBM neurospheres treated for 48 hours with 20 nM or 500 nM of bafilomycin A1, vehicle (Ctrl), or left untreated (Untr). Each point represents the mean of four experiments. RQ, mRNA Relative Quantity. D. <t>V-ATPase</t> G1 protein levels in GBM neurospheres after bafilomycin A1 treatment were analyzed by western blot. Vinculin was a loading control.
    Antibodies Against V Atpase G1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology v atpase d1
    YAP is highly expressed in EOC and is directly regulated by <t>V-ATPase</t> D1 expression. Immunohistochemical observation of V-ATPase D1 and YAP protein expression in normal ovarian epithelium and EOC tissues, a V-ATPase D1 and b YAP (with extremely weak staining (0), a weak staining (+ 1), a moderate staining (+ 2), a strong staining (+ 3). All photographs were taken at original 200 × magnification. The scores of ( a ) V-ATPase D1 and ( b ) YAP by beeswarm plot was shown in the bottom (** P
    V Atpase D1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology v atpase
    Quantitative analyses of the expression levels of <t>CD147</t> and <t>V-ATPase</t> in breast cancer samples. Comparison of the expression levels of (A) CD147 and (B) V-ATPase between the chemotherapy-sensitive and -resistant groups. (C) Correlation between the H-scores of CD147 and V-ATPase in breast invasive ductal cancer. ***P
    V Atpase, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti v atpase d2
    Astilbin represses NFATc1 activity and downstream protein expression. A, The bar graph depicts the NFATc1 luciferase activity of RAW264.7 cells stably transfected with an NFATc1 luciferase reporter construct. Cells were treated with varying concentrations of astilbin and stimulated by GST‐rRANKL (50 ng/mL) for 24 h. B, The protein expression of NFATc1, c‐Fos, <t>V‐ATPase‐d2,</t> CTSK and integrin β3 at day 0, day 1, day 3 and day 5 after stimulation by GST‐rRANKL (50 ng/mL) with or without astilbin (10 µmol/L). C‐G, Analysis of the statistical significance of the difference in protein expression between the astilbin‐treated group and control group. The expression of all proteins mentioned above was determined relative to β‐actin expression. The data in the figures represent means ± SD. Significant differences between the treatment and control groups are indicated as * P
    Anti V Atpase D2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Bafilomycin A1 repress stemness features of GBM neurospheres A. Nestin expression was evaluated by confocal immunofluorescence in GBM neurospheres incubated for 48 hours with vehicle or different concentrations of bafilomycin A1 and quantified in multiple z-stacks by ROI. **, p = 0.007; ***, p = 0.0006 by Mann-Whitney U test. Bars, mean ±SEM ( n = 5). Left , Representative maximum intensity projection images of neurospheres stained for the indicated antibody. B. , C. Gene expression evaluation of ATP6V1G1, CD133 and NESTIN, or of transcription factors involved in neurodevelopment and reprogramming (SOX2, OLIG2, SALL2 and POU3F2) in GBM neurospheres treated for 48 hours with 20 nM or 500 nM of bafilomycin A1, vehicle (Ctrl), or left untreated (Untr). Each point represents the mean of four experiments. RQ, mRNA Relative Quantity. D. V-ATPase G1 protein levels in GBM neurospheres after bafilomycin A1 treatment were analyzed by western blot. Vinculin was a loading control.

    Journal: Oncotarget

    Article Title: The vacuolar H+ ATPase is a novel therapeutic target for glioblastoma

    doi:

    Figure Lengend Snippet: Bafilomycin A1 repress stemness features of GBM neurospheres A. Nestin expression was evaluated by confocal immunofluorescence in GBM neurospheres incubated for 48 hours with vehicle or different concentrations of bafilomycin A1 and quantified in multiple z-stacks by ROI. **, p = 0.007; ***, p = 0.0006 by Mann-Whitney U test. Bars, mean ±SEM ( n = 5). Left , Representative maximum intensity projection images of neurospheres stained for the indicated antibody. B. , C. Gene expression evaluation of ATP6V1G1, CD133 and NESTIN, or of transcription factors involved in neurodevelopment and reprogramming (SOX2, OLIG2, SALL2 and POU3F2) in GBM neurospheres treated for 48 hours with 20 nM or 500 nM of bafilomycin A1, vehicle (Ctrl), or left untreated (Untr). Each point represents the mean of four experiments. RQ, mRNA Relative Quantity. D. V-ATPase G1 protein levels in GBM neurospheres after bafilomycin A1 treatment were analyzed by western blot. Vinculin was a loading control.

    Article Snippet: Glioma cultures were stained with primary antibodies against V-ATPase G1 (D5, Santa Cruz Biotechnologies), GFAP (G9269; Sigma Aldrich), cleaved Caspase-3 (D175; Cell Signaling Technologies, Danvers, MA, USA), Nestin (MAB 1259; R & D Systems, Abingdon, UK), Sox2 (D6D9, Cell Signaling Technology Inc.), Oligodendrocyte marker O4 (O7139; Sigma Aldrich), GFAP (G9269; Sigma Aldrich), or Tuji1 (T3952; Sigma Aldrich).

    Techniques: Expressing, Immunofluorescence, Incubation, MANN-WHITNEY, Staining, Western Blot

    Bafilomycin A1 reduces stem cell factors expression in GBM organ cultures A. Nestin immunoreactivity was analyzed in GBMs of the tissue micro-arrays and correlated to V-ATPase G1 expression in the same samples. B. , C. Short term GBM organ cultures were treated as in Figure 6 . Nestin presence was assessed after 72 hours and quantified relative to uncultured samples (T0). Original magnification x400. Bars, mean±SEM. §, p = 0.018 by Mann-Whitney U test. D. , E. The stem cell markers CD133, NESTIN or ATP6V1G1 ( D. ) and the neurodevelopmental transcription factors SOX2, OLIG2, SALL2 and POU3F2 ( E. ) were analyzed at gene expression level by qPCR in GBM organ cultures treated as indicated. Each point represents the mean of three cultures. RQ, mRNA Relative Quantity.

    Journal: Oncotarget

    Article Title: The vacuolar H+ ATPase is a novel therapeutic target for glioblastoma

    doi:

    Figure Lengend Snippet: Bafilomycin A1 reduces stem cell factors expression in GBM organ cultures A. Nestin immunoreactivity was analyzed in GBMs of the tissue micro-arrays and correlated to V-ATPase G1 expression in the same samples. B. , C. Short term GBM organ cultures were treated as in Figure 6 . Nestin presence was assessed after 72 hours and quantified relative to uncultured samples (T0). Original magnification x400. Bars, mean±SEM. §, p = 0.018 by Mann-Whitney U test. D. , E. The stem cell markers CD133, NESTIN or ATP6V1G1 ( D. ) and the neurodevelopmental transcription factors SOX2, OLIG2, SALL2 and POU3F2 ( E. ) were analyzed at gene expression level by qPCR in GBM organ cultures treated as indicated. Each point represents the mean of three cultures. RQ, mRNA Relative Quantity.

    Article Snippet: Glioma cultures were stained with primary antibodies against V-ATPase G1 (D5, Santa Cruz Biotechnologies), GFAP (G9269; Sigma Aldrich), cleaved Caspase-3 (D175; Cell Signaling Technologies, Danvers, MA, USA), Nestin (MAB 1259; R & D Systems, Abingdon, UK), Sox2 (D6D9, Cell Signaling Technology Inc.), Oligodendrocyte marker O4 (O7139; Sigma Aldrich), GFAP (G9269; Sigma Aldrich), or Tuji1 (T3952; Sigma Aldrich).

    Techniques: Expressing, MANN-WHITNEY, Real-time Polymerase Chain Reaction

    V-ATPase subunit G1 is a novel marker of poor prognosis in GBM patients A. , B. A glioma tissue microarray comprising 85 cores of normal brain parenchyma was analyzed for V-ATPase G1 expression by immunohistochemistry (IHC) A. Photomicrographs representing V-ATPase G1 immunoreactivity in normal brain tissue or in the indicated glioma histotype. Arrowheads indicate brain endothelium. B. V-ATPase G1 IHC score was obtained by multiplying the percentage of positive cells (range 0-100%) and the staining intensity (range: 0-3). According to this score, normal brain was almost negative (median IHC score 0; range: 0-10). Oligodendrogliomas (OD; median IHC score 1.75; range: 0-62.5), anaplastic OD (AO; median IHC score 6.1; range: 1.5-67.5), astrocytomas (Astr; median IHC score 6.2; range: 0.25-85), anaplastic astrocytomas (AA; median IHC score 11.75; range: 1-185) displayed higher V-ATPase G1 expression compared to normal brain. # , p = 0.0004; § , p = 0.047; °, p = 0.038; §§ , p = 0.005; §§§ , p

    Journal: Oncotarget

    Article Title: The vacuolar H+ ATPase is a novel therapeutic target for glioblastoma

    doi:

    Figure Lengend Snippet: V-ATPase subunit G1 is a novel marker of poor prognosis in GBM patients A. , B. A glioma tissue microarray comprising 85 cores of normal brain parenchyma was analyzed for V-ATPase G1 expression by immunohistochemistry (IHC) A. Photomicrographs representing V-ATPase G1 immunoreactivity in normal brain tissue or in the indicated glioma histotype. Arrowheads indicate brain endothelium. B. V-ATPase G1 IHC score was obtained by multiplying the percentage of positive cells (range 0-100%) and the staining intensity (range: 0-3). According to this score, normal brain was almost negative (median IHC score 0; range: 0-10). Oligodendrogliomas (OD; median IHC score 1.75; range: 0-62.5), anaplastic OD (AO; median IHC score 6.1; range: 1.5-67.5), astrocytomas (Astr; median IHC score 6.2; range: 0.25-85), anaplastic astrocytomas (AA; median IHC score 11.75; range: 1-185) displayed higher V-ATPase G1 expression compared to normal brain. # , p = 0.0004; § , p = 0.047; °, p = 0.038; §§ , p = 0.005; §§§ , p

    Article Snippet: Glioma cultures were stained with primary antibodies against V-ATPase G1 (D5, Santa Cruz Biotechnologies), GFAP (G9269; Sigma Aldrich), cleaved Caspase-3 (D175; Cell Signaling Technologies, Danvers, MA, USA), Nestin (MAB 1259; R & D Systems, Abingdon, UK), Sox2 (D6D9, Cell Signaling Technology Inc.), Oligodendrocyte marker O4 (O7139; Sigma Aldrich), GFAP (G9269; Sigma Aldrich), or Tuji1 (T3952; Sigma Aldrich).

    Techniques: Marker, Microarray, Expressing, Immunohistochemistry, Staining

    V-ATPase G1 levels are associated to clonogenicity of GBM neurospheres A. The ability of GBM cultures to generate neurospheres after specific ATP6V1G1 knockdown (V1G1-siRNA or esiRNA) relative to control sample was evaluated by time lapse microscopy for 72 hours. Representative images captured from movies ( Supplementary Movies 1 - 3 ) at time zero and at harvesting are shown. B. GBM neurospheres clonogenic capacity in methylcellulose containing media was analyzed in the indicated samples. Left , quantification of spheres containing at least 20 cells. C. V-ATPase G1 gene or protein expression levels in the indicated GBM cultures. D. The clonogenic potential of the indicated GBM neurosphere was assessed after V1G1 knockdown by siRNA or esiRNA and compared to corresponding controls. *, p

    Journal: Oncotarget

    Article Title: The vacuolar H+ ATPase is a novel therapeutic target for glioblastoma

    doi:

    Figure Lengend Snippet: V-ATPase G1 levels are associated to clonogenicity of GBM neurospheres A. The ability of GBM cultures to generate neurospheres after specific ATP6V1G1 knockdown (V1G1-siRNA or esiRNA) relative to control sample was evaluated by time lapse microscopy for 72 hours. Representative images captured from movies ( Supplementary Movies 1 - 3 ) at time zero and at harvesting are shown. B. GBM neurospheres clonogenic capacity in methylcellulose containing media was analyzed in the indicated samples. Left , quantification of spheres containing at least 20 cells. C. V-ATPase G1 gene or protein expression levels in the indicated GBM cultures. D. The clonogenic potential of the indicated GBM neurosphere was assessed after V1G1 knockdown by siRNA or esiRNA and compared to corresponding controls. *, p

    Article Snippet: Glioma cultures were stained with primary antibodies against V-ATPase G1 (D5, Santa Cruz Biotechnologies), GFAP (G9269; Sigma Aldrich), cleaved Caspase-3 (D175; Cell Signaling Technologies, Danvers, MA, USA), Nestin (MAB 1259; R & D Systems, Abingdon, UK), Sox2 (D6D9, Cell Signaling Technology Inc.), Oligodendrocyte marker O4 (O7139; Sigma Aldrich), GFAP (G9269; Sigma Aldrich), or Tuji1 (T3952; Sigma Aldrich).

    Techniques: esiRNA, Time-lapse Microscopy, Expressing

    YAP is highly expressed in EOC and is directly regulated by V-ATPase D1 expression. Immunohistochemical observation of V-ATPase D1 and YAP protein expression in normal ovarian epithelium and EOC tissues, a V-ATPase D1 and b YAP (with extremely weak staining (0), a weak staining (+ 1), a moderate staining (+ 2), a strong staining (+ 3). All photographs were taken at original 200 × magnification. The scores of ( a ) V-ATPase D1 and ( b ) YAP by beeswarm plot was shown in the bottom (** P

    Journal: BMC Molecular and Cell Biology

    Article Title: Proton pump inhibitors can reverse the YAP mediated paclitaxel resistance in epithelial ovarian cancer

    doi: 10.1186/s12860-019-0227-y

    Figure Lengend Snippet: YAP is highly expressed in EOC and is directly regulated by V-ATPase D1 expression. Immunohistochemical observation of V-ATPase D1 and YAP protein expression in normal ovarian epithelium and EOC tissues, a V-ATPase D1 and b YAP (with extremely weak staining (0), a weak staining (+ 1), a moderate staining (+ 2), a strong staining (+ 3). All photographs were taken at original 200 × magnification. The scores of ( a ) V-ATPase D1 and ( b ) YAP by beeswarm plot was shown in the bottom (** P

    Article Snippet: Statistical analysis The correlation between YAP and V-ATPase D1 was evaluated using Spearman’s rank correlation coefficient test.

    Techniques: Expressing, Immunohistochemistry, Staining

    Effects of acidic pHe on V-ATPase D1 change. a Total protein from PTX- resistant EOC cells and respective sensitive phenotypes were immuno-blotted with anti-V-ATPase D1. b Confocal microscopy showing V-ATPase D1 (green dots) and plasma membrane (red/orange staining) in A2780 and A2780/T cells. DAPI in blue color represents nuclear staining. Merged images (yellow regions). Original magnification× 600. Representative images from three independent experiments are shown. c Cytosolic pH was estimated using pH sensitive dye BCECF-AM. Significantly decreased intracellular pH was observed in EMSO co-treated with PTX in A2780/T cells using BCECF-AM and fluorescence microscopy (488/535 nm, Magnification× 600). d The corresponding intracellular pH was obtained from the pH calibration curve. Values are means ± S.E.M of two independent experiments performed in triplicate (* P

    Journal: BMC Molecular and Cell Biology

    Article Title: Proton pump inhibitors can reverse the YAP mediated paclitaxel resistance in epithelial ovarian cancer

    doi: 10.1186/s12860-019-0227-y

    Figure Lengend Snippet: Effects of acidic pHe on V-ATPase D1 change. a Total protein from PTX- resistant EOC cells and respective sensitive phenotypes were immuno-blotted with anti-V-ATPase D1. b Confocal microscopy showing V-ATPase D1 (green dots) and plasma membrane (red/orange staining) in A2780 and A2780/T cells. DAPI in blue color represents nuclear staining. Merged images (yellow regions). Original magnification× 600. Representative images from three independent experiments are shown. c Cytosolic pH was estimated using pH sensitive dye BCECF-AM. Significantly decreased intracellular pH was observed in EMSO co-treated with PTX in A2780/T cells using BCECF-AM and fluorescence microscopy (488/535 nm, Magnification× 600). d The corresponding intracellular pH was obtained from the pH calibration curve. Values are means ± S.E.M of two independent experiments performed in triplicate (* P

    Article Snippet: Statistical analysis The correlation between YAP and V-ATPase D1 was evaluated using Spearman’s rank correlation coefficient test.

    Techniques: Confocal Microscopy, Staining, Fluorescence, Microscopy

    Quantitative analyses of the expression levels of CD147 and V-ATPase in breast cancer samples. Comparison of the expression levels of (A) CD147 and (B) V-ATPase between the chemotherapy-sensitive and -resistant groups. (C) Correlation between the H-scores of CD147 and V-ATPase in breast invasive ductal cancer. ***P

    Journal: Oncology Letters

    Article Title: Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity

    doi: 10.3892/ol.2018.8199

    Figure Lengend Snippet: Quantitative analyses of the expression levels of CD147 and V-ATPase in breast cancer samples. Comparison of the expression levels of (A) CD147 and (B) V-ATPase between the chemotherapy-sensitive and -resistant groups. (C) Correlation between the H-scores of CD147 and V-ATPase in breast invasive ductal cancer. ***P

    Article Snippet: Subsequent to washing with PBS, the cells were incubated for 2 h at room temperature with primary antibodies against CD147 (1:200; cat. no. ab212856; Abcam) and V-ATPase (1:100; cat. no. sc-69088; Santa Cruz Biotechnology, Inc. Dallas, TX, USA), followed by incubation with secondary antibodies, including anti-mouse Cy3 (1:200; cat. no. AP192C; EMD Millipore, Billerica, MA, USA) or anti-goat FITC (1:500; cat. no. ab6881; Abcam) antibodies for 1 h at room temperature.

    Techniques: Expressing

    Immunohistochemical observation of CD147 and V-ATPase expression in chemotherapy-sensitive or -resistant breast invasive ductal cancer patients (magnification, ×400). Chemotherapy-sensitive patients exhibited moderate expression of (A) CD147 and (B) V-ATPase in the cell membrane. Chemotherapy-resistant patients exhibited high expression of (C) CD147 in the cell membrane and (D) V-ATPase mainly in the cell membrane and cytoplasm. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase.

    Journal: Oncology Letters

    Article Title: Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity

    doi: 10.3892/ol.2018.8199

    Figure Lengend Snippet: Immunohistochemical observation of CD147 and V-ATPase expression in chemotherapy-sensitive or -resistant breast invasive ductal cancer patients (magnification, ×400). Chemotherapy-sensitive patients exhibited moderate expression of (A) CD147 and (B) V-ATPase in the cell membrane. Chemotherapy-resistant patients exhibited high expression of (C) CD147 in the cell membrane and (D) V-ATPase mainly in the cell membrane and cytoplasm. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase.

    Article Snippet: Subsequent to washing with PBS, the cells were incubated for 2 h at room temperature with primary antibodies against CD147 (1:200; cat. no. ab212856; Abcam) and V-ATPase (1:100; cat. no. sc-69088; Santa Cruz Biotechnology, Inc. Dallas, TX, USA), followed by incubation with secondary antibodies, including anti-mouse Cy3 (1:200; cat. no. AP192C; EMD Millipore, Billerica, MA, USA) or anti-goat FITC (1:500; cat. no. ab6881; Abcam) antibodies for 1 h at room temperature.

    Techniques: Immunohistochemistry, Expressing

    Immunofluorescence was used to detect CD147 and V-ATPase expression and subcellular localization in the four transfected cell lines. Experiments were repeated at least three times, and a representative experiment is shown. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase; CON, control; CD, CD147 overexpression; si, CD147 knockdown. Blue represents nuclear staining; red represents CD147 staining; yellow represents V-ATPase staining.

    Journal: Oncology Letters

    Article Title: Cluster of differentiation 147 mediates chemoresistance in breast cancer by affecting vacuolar H+-ATPase expression and activity

    doi: 10.3892/ol.2018.8199

    Figure Lengend Snippet: Immunofluorescence was used to detect CD147 and V-ATPase expression and subcellular localization in the four transfected cell lines. Experiments were repeated at least three times, and a representative experiment is shown. CD147, cluster of differentiation 147; V-ATPase, vacuolar H + -ATPase; CON, control; CD, CD147 overexpression; si, CD147 knockdown. Blue represents nuclear staining; red represents CD147 staining; yellow represents V-ATPase staining.

    Article Snippet: Subsequent to washing with PBS, the cells were incubated for 2 h at room temperature with primary antibodies against CD147 (1:200; cat. no. ab212856; Abcam) and V-ATPase (1:100; cat. no. sc-69088; Santa Cruz Biotechnology, Inc. Dallas, TX, USA), followed by incubation with secondary antibodies, including anti-mouse Cy3 (1:200; cat. no. AP192C; EMD Millipore, Billerica, MA, USA) or anti-goat FITC (1:500; cat. no. ab6881; Abcam) antibodies for 1 h at room temperature.

    Techniques: Immunofluorescence, Expressing, Transfection, Over Expression, Staining

    Astilbin represses NFATc1 activity and downstream protein expression. A, The bar graph depicts the NFATc1 luciferase activity of RAW264.7 cells stably transfected with an NFATc1 luciferase reporter construct. Cells were treated with varying concentrations of astilbin and stimulated by GST‐rRANKL (50 ng/mL) for 24 h. B, The protein expression of NFATc1, c‐Fos, V‐ATPase‐d2, CTSK and integrin β3 at day 0, day 1, day 3 and day 5 after stimulation by GST‐rRANKL (50 ng/mL) with or without astilbin (10 µmol/L). C‐G, Analysis of the statistical significance of the difference in protein expression between the astilbin‐treated group and control group. The expression of all proteins mentioned above was determined relative to β‐actin expression. The data in the figures represent means ± SD. Significant differences between the treatment and control groups are indicated as * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Astilbin prevents bone loss in ovariectomized mice through the inhibition of RANKL‐induced osteoclastogenesis, et al. Astilbin prevents bone loss in ovariectomized mice through the inhibition of RANKL‐induced osteoclastogenesis

    doi: 10.1111/jcmm.14713

    Figure Lengend Snippet: Astilbin represses NFATc1 activity and downstream protein expression. A, The bar graph depicts the NFATc1 luciferase activity of RAW264.7 cells stably transfected with an NFATc1 luciferase reporter construct. Cells were treated with varying concentrations of astilbin and stimulated by GST‐rRANKL (50 ng/mL) for 24 h. B, The protein expression of NFATc1, c‐Fos, V‐ATPase‐d2, CTSK and integrin β3 at day 0, day 1, day 3 and day 5 after stimulation by GST‐rRANKL (50 ng/mL) with or without astilbin (10 µmol/L). C‐G, Analysis of the statistical significance of the difference in protein expression between the astilbin‐treated group and control group. The expression of all proteins mentioned above was determined relative to β‐actin expression. The data in the figures represent means ± SD. Significant differences between the treatment and control groups are indicated as * P

    Article Snippet: The membranes were incubated overnight at 4°C with the following primary antibodies: anti‐NFATc1 (1:1000; Santa Cruz Biotechnology), anti‐c‐Fos (1:2000; Santa Cruz Biotechnology), anti‐integrin β3 (1:1000; Santa Cruz Biotechnology), anti‐CTSK (1:2000, Santa Cruz Biotechnology), anti‐V‐ATPase‐d2 (1:1000; Santa Cruz Biotechnology), anti‐IκB‐α (1:1000; Santa Cruz Biotechnology), anti‐p‐ERK1/2 (1:1000, Santa Cruz Biotechnology), anti‐ERK1/2 (1:1000; Santa Cruz Biotechnology), anti‐p‐JNK1/2 (1:1000; Cell Signaling Technologies), anti‐JNK1/2 (1:5000, 1:1000; Cell Signaling Technologies), anti‐p‐p38 (1:1000; Cell Signaling Technologies), anti‐p38 (1:1000; Cell Signaling Technologies) and anti‐β‐actin (1:3000; Cell Signaling Technologies); later, the membranes were immersed for 1 hour in corresponding horseradish peroxidase‐conjugated secondary antibodies.

    Techniques: Activity Assay, Expressing, Luciferase, Stable Transfection, Transfection, Construct