Journal: Oncology Letters

Article Title: Association between Twist and multidrug resistance gene-associated proteins in Taxol ® -resistant MCF-7 cells and a 293 cell model of Twist overexpression

doi: 10.3892/ol.2017.7438

Figure Lengend Snippet: Expression of Twist and drug resistance-associated proteins in human breast cancer. The immunohistochemistry results (magnification, ×400) demonstrated that, compared with the pericancerous tissue, the expression of Twist, TOPO IIα, MRP and P-gp were upregulated in cancerous tissues, whereas the expression of LRP was downregulated in cancerous tissues. TOPO IIα, topoisomerase IIα; MRP, multidrug resistance-associated protein; P-gp, P-glycoprotein.

Article Snippet: The primary antibodies used in the present study included: Mouse anti-TOPO IIα antibody (dilution, 1:200; cat. no. ZM-0245; OriGene Technologies, Inc.); mouse anti-MRP antibody (dilution, 1:200; cat. no. ZM-0345; OriGene Technologies, Inc.); rabbit anti-P-gp antibody (dilution, 1:500; cat. no. ab103477; Abcam, Cambridge, UK); mouse anti-LRP antibody (dilution, 1:200; cat. no. ZM-0325; OriGene Technologies, Inc.); and rabbit anti-Twist antibody (dilution, 1:500; cat. no. ab49254; Abcam).

Techniques: Expressing, Immunohistochemistry

Journal: Oncology Letters

Article Title: Association between Twist and multidrug resistance gene-associated proteins in Taxol ® -resistant MCF-7 cells and a 293 cell model of Twist overexpression

doi: 10.3892/ol.2017.7438

Figure Lengend Snippet: Taxol ® resistance increases the protein expression of Twist, TOPO IIα, MRP and P-gp in MCF-7/Taxol ® cells. Immunohistochemistry results (magnification, ×400) demonstrated that the protein expression of Twist, TOPO IIα, MRP and P-gp were increased in Taxol ® -resistant cells, whereas the protein expression of LRP was reduced. TOPO IIα, topoisomerase IIα; MRP, multidrug resistant-associated protein; P-gp, P-glycoprotein; LRP, lung resistance-related protein.

Article Snippet: The primary antibodies used in the present study included: Mouse anti-TOPO IIα antibody (dilution, 1:200; cat. no. ZM-0245; OriGene Technologies, Inc.); mouse anti-MRP antibody (dilution, 1:200; cat. no. ZM-0345; OriGene Technologies, Inc.); rabbit anti-P-gp antibody (dilution, 1:500; cat. no. ab103477; Abcam, Cambridge, UK); mouse anti-LRP antibody (dilution, 1:200; cat. no. ZM-0325; OriGene Technologies, Inc.); and rabbit anti-Twist antibody (dilution, 1:500; cat. no. ab49254; Abcam).

Techniques: Expressing, Immunohistochemistry

Journal: Oncology Letters

Article Title: Association between Twist and multidrug resistance gene-associated proteins in Taxol ® -resistant MCF-7 cells and a 293 cell model of Twist overexpression

doi: 10.3892/ol.2017.7438

Figure Lengend Snippet: Immunohistochemistry analysis of LRP, TOPO IIα, MRP, P-gp and Twist in Taxol-resistant breast cancer cell lines.

Article Snippet: The primary antibodies used in the present study included: Mouse anti-TOPO IIα antibody (dilution, 1:200; cat. no. ZM-0245; OriGene Technologies, Inc.); mouse anti-MRP antibody (dilution, 1:200; cat. no. ZM-0345; OriGene Technologies, Inc.); rabbit anti-P-gp antibody (dilution, 1:500; cat. no. ab103477; Abcam, Cambridge, UK); mouse anti-LRP antibody (dilution, 1:200; cat. no. ZM-0325; OriGene Technologies, Inc.); and rabbit anti-Twist antibody (dilution, 1:500; cat. no. ab49254; Abcam).

Techniques: Immunohistochemistry

Journal: Oncology Letters

Article Title: Association between Twist and multidrug resistance gene-associated proteins in Taxol ® -resistant MCF-7 cells and a 293 cell model of Twist overexpression

doi: 10.3892/ol.2017.7438

Figure Lengend Snippet: Overexpression of Twist regulates the expression of multidrug resistance-related proteins in 293 cells. The immunohistochemistry results (magnification, ×400) indicated that the protein expression of Twist and P-gp were elevated in 293-Twist cells. However, no difference in protein expression of LRP, TOPO IIα and MRP were observed in 293-Twist cells and 293-vector cells. TOPO IIα, topoisomerase IIα; MRP, multidrug resistant-associated protein; P-gp, P-glycoprotein; LRP, lung resistance-related protein.

Article Snippet: The primary antibodies used in the present study included: Mouse anti-TOPO IIα antibody (dilution, 1:200; cat. no. ZM-0245; OriGene Technologies, Inc.); mouse anti-MRP antibody (dilution, 1:200; cat. no. ZM-0345; OriGene Technologies, Inc.); rabbit anti-P-gp antibody (dilution, 1:500; cat. no. ab103477; Abcam, Cambridge, UK); mouse anti-LRP antibody (dilution, 1:200; cat. no. ZM-0325; OriGene Technologies, Inc.); and rabbit anti-Twist antibody (dilution, 1:500; cat. no. ab49254; Abcam).

Techniques: Over Expression, Expressing, Immunohistochemistry, Plasmid Preparation

Journal: Pharmaceutical research

Article Title: In Vitro Transport Activity and Trafficking of MRP2/ABCC2 Polymorphic Variants.

doi: 10.1007/s11095-017-2160-0

Figure Lengend Snippet: Fig. 1 Localization of variants in a predicted topology model of the human MRP2 transporter. The full-length model of the MRP2 protein was generated using the open source tool Protter (http://wlab.ethz.ch/protter/start/). The C-24T variant is found in the 5′-untranslated region of the ABCC2 promoter and not shown in the figure. MSD: membrane-spanning domain; NBD: nucleotide-binding domain.

Article Snippet: The full-length human ABCC2WTcDNA clone was obtained by digestion of a pCMV6-NEO plasmid containing ABCC2 cDNA (Origene, Rockville, MD) using the restriction enzymes SalI and NotI (New England Biolabs, Ipswich, MA).

Techniques: Generated, Variant Assay, Membrane, Binding Assay

Journal: Pharmaceutical research

Article Title: In Vitro Transport Activity and Trafficking of MRP2/ABCC2 Polymorphic Variants.

doi: 10.1007/s11095-017-2160-0

Figure Lengend Snippet: Fig. 2 Expression and activity of the MRP2 transporter using the Flp-In system. Flp-In 293 cells were stably transfected with empty vector (EV) or full-length human ABCC2 plasmids (wild-type, WT). Protein expression of MRP2 in cell lysates (a) and biotinylated membrane fractions (b) was detected by Western blot analysis. β-ACTIN and Na+/K+ ATPase were used as loading controls for total and cell surface expression, respectively. (c) EV and WT-MRP2 cells were treated with calcein AM (0.5 μM) for 30 min (uptake period), washed, and then incubated with fresh culture media for 60 min (efflux period). Intracellular accumulation of calcein AM was determined at the end of the efflux period using a fluorescent cell counter. Data are expressed as relative fluorescence units (RFU) and presented as mean ± SE (n = 4). Asterisks (*) represent statistically significant differences (p < 0.05) compared to EV.

Article Snippet: The full-length human ABCC2WTcDNA clone was obtained by digestion of a pCMV6-NEO plasmid containing ABCC2 cDNA (Origene, Rockville, MD) using the restriction enzymes SalI and NotI (New England Biolabs, Ipswich, MA).

Techniques: Expressing, Activity Assay, Stable Transfection, Transfection, Plasmid Preparation, Membrane, Western Blot, Incubation, Fluorescence

Journal: Pharmaceutical research

Article Title: In Vitro Transport Activity and Trafficking of MRP2/ABCC2 Polymorphic Variants.

doi: 10.1007/s11095-017-2160-0

Figure Lengend Snippet: Fig. 3 Transport of calcein AM in MRP2 wild-type and variant cells. Flp-In 293 cells stably expressing ABCC2 wild-type (WT) or variant plasmids were exposed to calcein AM for 30 min (uptake period), washed, and then incubated with fresh culture medium for 60 min (efflux period). Intracellular calcein AM accumulation was determined at the end of the efflux period using a fluorescent counter. (a) WT-MRP2 and variant cells were exposed to calcein AM (0.5 μM). (b) WT-MRP2, C2366T, G3542 Tand C3972T variants were exposed to calcein AM (0.5, 2, and 10 μM). Data are expressed as relative fluorescence units (RFU) and presented as mean ± SE (n = 4–6). Asterisks (*) represent statistically significant differences (p < 0.05) compared to WT.

Article Snippet: The full-length human ABCC2WTcDNA clone was obtained by digestion of a pCMV6-NEO plasmid containing ABCC2 cDNA (Origene, Rockville, MD) using the restriction enzymes SalI and NotI (New England Biolabs, Ipswich, MA).

Techniques: Variant Assay, Stable Transfection, Expressing, Incubation, Fluorescence

Journal: Pharmaceutical research

Article Title: In Vitro Transport Activity and Trafficking of MRP2/ABCC2 Polymorphic Variants.

doi: 10.1007/s11095-017-2160-0

Figure Lengend Snippet: Fig. 4 Protein expression and localization of MRP2 in wild-type and variant cells. Flp-In 293 cells were stably transfected with ABCC2 wild-type (WT) or variant plasmids. Protein expression of MRP2 in cell lysates (a) and biotinylated membrane fractions (b) was detected by Western blot analysis. β-ACTIN and Na+/K+ ATPase were used as loading controls for total and cell surface expression, respectively. Data are expressed as relative protein expression and presented as mean ± SE (n = 3). Asterisks (*) represent statistically significant differences (p < 0.05) compared to WT.

Article Snippet: The full-length human ABCC2WTcDNA clone was obtained by digestion of a pCMV6-NEO plasmid containing ABCC2 cDNA (Origene, Rockville, MD) using the restriction enzymes SalI and NotI (New England Biolabs, Ipswich, MA).

Techniques: Expressing, Variant Assay, Stable Transfection, Transfection, Membrane, Western Blot

Journal: Pharmaceutical research

Article Title: In Vitro Transport Activity and Trafficking of MRP2/ABCC2 Polymorphic Variants.

doi: 10.1007/s11095-017-2160-0

Figure Lengend Snippet: Fig. 5 Localization of MRP2 protein in wild-type and variant cells. Flp-In 293 cells were stably transfected with ABCC2 wild-type (WT) or variant plasmids. Staining of MRP2 protein in WT, G1249A, C2366T, G3542T, T3563A, C3972T, and G4544A variant cells was performed using indirect immunofluorescence analysis. Cells were incubated with an anti-MRP2 antibody (M2III-6) (green). Images are shown at 63X magnification.

Article Snippet: The full-length human ABCC2WTcDNA clone was obtained by digestion of a pCMV6-NEO plasmid containing ABCC2 cDNA (Origene, Rockville, MD) using the restriction enzymes SalI and NotI (New England Biolabs, Ipswich, MA).

Techniques: Variant Assay, Stable Transfection, Transfection, Staining, Immunofluorescence, Incubation

Journal: Pharmaceutical research

Article Title: In Vitro Transport Activity and Trafficking of MRP2/ABCC2 Polymorphic Variants.

doi: 10.1007/s11095-017-2160-0

Figure Lengend Snippet: Fig. 6 Expression of MRP2 protein and CDCF transport in membrane vesicles isolated from wild-type and variant cells. Flp-In 293 cells were stably transfected with ABCC2 wild-type (WT) or variant plasmids. (a) Protein expression of MRP2 in membrane vesicles isolated from cells was detected by Western blot analysis. Na+/K+ ATPase was used as a loading control. (b) CDCF (10 μM) transport in membrane vesicles isolated from cells expressing WT-MRP2 and variants. Transport of CDCF (10 μM) in empty vector-transfected vesicles was 4.2 pmol/min/mg protein. (c) CDCF (2.5, 10 and 50 μM) transport in membrane vesicles isolated from cells expressing WT-MRP2 and variants G1249A, C2366T, G3542 T. Transport was conducted using vesicles (50 μg) incubated with CDCF at 37°C for 10 min. Data are presented as mean ± SE (n = 3–6). Asterisks (*) represent statistically significant differences (p < 0.05) compared to WT.

Article Snippet: The full-length human ABCC2WTcDNA clone was obtained by digestion of a pCMV6-NEO plasmid containing ABCC2 cDNA (Origene, Rockville, MD) using the restriction enzymes SalI and NotI (New England Biolabs, Ipswich, MA).

Techniques: Expressing, Membrane, Isolation, Variant Assay, Stable Transfection, Transfection, Western Blot, Control, Plasmid Preparation, Incubation

Journal: Neurotoxicology

Article Title: Contribution of child ABC-transporter genetics to prenatal MeHg exposure and neurodevelopment

doi: 10.1016/j.neuro.2022.05.019

Figure Lengend Snippet: Summary of current knowledge and distribution in different populations for the six SNPs evaluated in the study.

Article Snippet: Genotyping was performed by TaqMan real-time PCR on the ABI 7900HT Fast Real Time PCR System (Applied Biosystems, Thermo Fisher, Waltham, USA), using manufacturer’s recommended standard conditions and the following custom genotyping assays from Thermo Scientific: C__11711720C_30 and C__11711720D_40 (rs2032582), C__29490882_10 (rs10276499), C__2982705_20 (rs1202169), C__3188828_10 (rs212093), C__27065543_10 (rs215088) C__2814642_10 (rs717620).

Techniques: Expressing, Membrane

Journal: Neurotoxicology

Article Title: Contribution of child ABC-transporter genetics to prenatal MeHg exposure and neurodevelopment

doi: 10.1016/j.neuro.2022.05.019

Figure Lengend Snippet: Mean cord blood (prenatal) MeHg concentrations by child and maternal genotypes of ABC transporter SNP. For each SNP, the p-value for the test comparing the means in the three groups is reported.

Article Snippet: Genotyping was performed by TaqMan real-time PCR on the ABI 7900HT Fast Real Time PCR System (Applied Biosystems, Thermo Fisher, Waltham, USA), using manufacturer’s recommended standard conditions and the following custom genotyping assays from Thermo Scientific: C__11711720C_30 and C__11711720D_40 (rs2032582), C__29490882_10 (rs10276499), C__2982705_20 (rs1202169), C__3188828_10 (rs212093), C__27065543_10 (rs215088) C__2814642_10 (rs717620).

Techniques: