Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Conditional deletion of Abcb5 in mice (A) Schematic representation of the strategy used to target the Abcb5 gene. LoxP sites (red triangles) were inserted upstream and downstream of exon 2 (E2). The neomycin resistance cassette ( NEO ) is flanked by Frt sites (yellow ovals). (B) After the initial screening of the positive ES cell clones, a 5′ external probe (red rectangle) was used to identify the targeted alleles by detecting a shift from the endogenous HindIII wild-type (WT) band of 13 kb to the rearranged 8.7 Kb band resulting from the insertion of the neo cassette by homologous recombination. The correct targeting was confirmed by using a 3′ external probe following digestion with the HindIII restriction enzyme, which detects a shift from the 13 Kb endogenous WT band to the rearranged 6.6 Kb mutant band. HSV-TK, thymidine kinase cassette for the negative selection; pBS, backbone pbluescript vector. (C) The presence of the distal loxP site in the genome was tested by PCR. Amplification of the WT or mutant Abcb5 alleles resulted in bands of either 239 bp (WT) or 330 bp (recombined allele). (D) Mice were routinely genotyped by isolating genomic DNA from an ear punch using 100 ng of genomic DNA as the template. Amplification of the WT or mutant Abcb5 alleles resulted in bands of either 512 bp (Abcb5 +/+ ) or 112 bp (Abcb5 −/− ).

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Clone Assay, Homologous Recombination, Mutagenesis, Selection, Plasmid Preparation, Amplification

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: The Abcb5-mutant mice have lower lactate levels, indicating a shift in cellular energy metabolism (A) Lactate levels [mmol/L] in female (f) and male (m) Abcb5 +/+ (control, con) and Abcb5 −/− (mutant, mut) mice. Reduced lactate levels were detected in the ABCB5 −/− mice ( p < 0.01), . (B) Fasting cholesterol boxplot with stripchart, split by sex and genotype ( p = 0.046), . (C) Fasting HDL cholesterol boxplot with stripchart, split by sex and genotype ( p = 0.028), . (D) Triglyceride boxplot with stripchart, split by sex and genotype ( p = 0.016), . Two cohorts, each consisting of 10 male and 10 female WT and homozygous mutant adult mice, all born within one week, were subjected to clinical chemistry analyses at 13 and 14 weeks of age. Means, standard deviations, and p -values for genotype, sex, and genotype–sex interaction effects were calculated using a linear model. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Mutagenesis, Control

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Lipid alteration of Abcb5-mutant mice heart and liver (A) The number of heart and liver lipid droplets after staining with Oil Red O ( n = 10). Droplets were counted using ImageJ. Abcb5-deficient mice showed a reduced number of lipid droplets in the heart ( p = 0.011), and a female-specific decrease in droplet number in the liver ( p = 0.021). (B) Diameter of heart and lipid droplets after staining with Oil Red O ( n = 10). The Droplets diameter was determined using ImageJ. A female-specific decrease in droplet size in the liver was observed ( p = 0.019). (C) Representative image of heart sections stained with Oil Red O. (D) Representative image of liver sections stained with Oil Red O. Two cohorts, each consisting of 10 male and 10 female WT and homozygous mutant adult mice, all born within one week, were analyzed for heart and liver lipid content at 8 weeks of age. Means, standard deviations, and p -values for genotype, sex, and genotype–sex interaction effects were calculated using a two-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Scale bar = 100 μM.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Mutagenesis, Staining

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Variation of ketone bodies levels in Abcb5-mutant mice Quantification of ketone bodies (acetoacetate, acetate, β-hydroxybutyrate) in the plasma (A), heart (B), and liver (C). Variations in acetoacetate levels were observed in the liver ( p = 0.028) of Abcb5-knockout mice. A female-specific decrease and a male-specific increase in β-hydroxybutyrate levels was observed in heart ( p = 0.048) and liver ( p = 0.020). A similar observation was made for acetate levels in the liver ( p = 0.028). Two cohorts, each consisting of 10 male and 10 female WT and homozygous mutant adult mice, all born within one week, were analyzed for ketone body levels at 8 weeks of age. Means, standard deviations, and p -values for genotype, sex, and genotype–sex interaction effects were calculated using a two-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. See .

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Mutagenesis, Clinical Proteomics, Knock-Out

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: ERα activates ABCB5β transcription MelJuSo were transfected with pSG5-hERα and either 3xERE::luc-TATA, ABCB5β_3xERE::luc-TATA or ABCB5β_0xERE::luc-TATA ( n = 6). 24 h post-transfection, cells were incubated with 5 mM estrogen. Then, luminescence was analyzed using the Dual-Luciferase Reporter Assay System (Promega). The y axis represents the luminescence fold change compared to MelJuSo transfected with pSG5 empty plasmid and 3xERE::luc-TATA in the absence of estrogen.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Transfection, Incubation, Luciferase, Reporter Assay, Plasmid Preparation

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Differential Abc gene expression profile in Abcb5-knockout mice Representation of the impact of Abcb5 KO on the expression of other Abc genes in seven different organs. Points and error bars represent geometric means and 95% confidence intervals on the mean, respectively. Statistical significance is represented using darker dots. The mRNA expression profile of 52 murine Abc transporters was measured in seven tissues collected from 5 male and 5 female WT and homozygous mutant adult mice, all born within one week, and analyzed at 12 weeks of age.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Gene Expression, Knock-Out, Expressing, Mutagenesis

Journal: iScience

Article Title: Abcb5-deficient mice show a subtle, pleiotropic phenotype indicating a role for this transporter in intermediary metabolism

doi: 10.1016/j.isci.2025.113617

Figure Lengend Snippet: Heatmap representing the impact of Abcb5 knockout on the expression of other Abc genes in seven different organs Tile colors represent the ΔΔCt values. Low ΔΔCt corresponds to overexpression and is red colored. Genes and organs are clustered following the Euclidean distance; data are not scaled. The mRNA expression profile of 52 murine ABC transporters was measured in seven tissues collected from 5 male and 5 female WT and homozygous mutant adult mice, all born within one week, and analyzed at 12 weeks of age.

Article Snippet: Abcb5 , ThermoFisher Scientific , Mm01225815_m1.

Techniques: Knock-Out, Expressing, Over Expression, Mutagenesis

Journal: Regenerative Therapy

Article Title: Conditioned media of stem cells from human exfoliated deciduous teeth contain factors related to extracellular matrix organization and promotes corneal epithelial wound healing

doi: 10.1016/j.reth.2025.03.002

Figure Lengend Snippet: Immunohistochemical findings of limbal stem cell markers in the limbal epithelium of chronic graft-versus-host disease (cGVHD) mice. Sections of the limbus from syngeneic control mice and cGVHD mice treated with vehicle, stem cells from human exfoliated deciduous teeth-conditioned medium (SHED-CM), immortalized-SHED-CM (IM-SHED-CM), or >3.5 kD fractionated components were stained for ATP-binding cassette sub-family B member 5 (ABCB5) (green) (a) or p63 (green) (b) (n = 5–6, per group). The corneal epithelium is located above the white dotted line. Data are presented as the mean ± standard error of the mean. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. One-way analysis of variance with Tukey–Kramer's post-hoc test was used for the analysis. Scale bar, 10 μm.

Article Snippet: This process was applied to antibodies against p63 (dilution of 1:500, sc-25268, monoclonal, Santa Cruz Biotechnology, Dallas, TX), ATP-binding cassette sub-family B member 5 (ABCB5) (dilution of 1:500, bs-1604R, Bioss Antibodies, Woburn, MA), 4-hydrogy-2-nonenal (4-HNE) (dilution of 1:100, HNEJ-2, Japan Institute for the Control Aging, Shizuoka, Japan), or 8-hydroxy-2′-deoxyguanosine (8-OHdG) (dilution of 1:50, MOG-020P, N45.1, Genox, Torrance, CA).

Techniques: Immunohistochemical staining, Control, Staining, Binding Assay

Journal: Current Issues in Molecular Biology

Article Title: Valproic Acid as a Histone Deacetylase Inhibitor Induces ABCB1 Overexpression and De Novo ABCB5 Expression in HeLa Cells

doi: 10.3390/cimb47090749

Figure Lengend Snippet: Protein sequence detected by Novus ABCB5 monoclonal antibody and its comparison to variants 1 and 2. The immunogen comprises 194 amino acids ( A ). ABCB5 protein immunodetection in HeLa cells treated with 5 mM VPA ( B ). NT: non-treated.

Article Snippet: The cells were fixed with 100% ice-cold methanol, blocked with 5% normal goat serum, and incubated overnight at 4 °C with a 1:200 dilution of primary ABCB5 monoclonal antibody (Novus Biologicals, Littleton, CO, USA).

Techniques: Sequencing, Comparison, Immunodetection

Journal: Oncology Research

Article Title: Identifying ATP-Binding Cassette Member B5 as a New Biomarker for Oral Squamous Cell Carcinoma

doi: 10.32604/or.2025.064276

Figure Lengend Snippet: ABCB5 is highly expressed in oral cancer cell lines: ( a–e ) The mRNA and protein expressions of ABCB5 were measured in HOK, CAL27, and HSC-3 using qRT-PCR, western blot, and ICC. ( a ) ABCB5 mRNA level in HOK, CAL27, and HSC-3 was detected by qRT-PCR, n = 3 qRT-PCR runs. ( b,c ) The ABCB5 protein level in HOK, CAL27, and HSC-3 was measured by western blot ( b ), the density of ABCB5 was quantified by ImageJ, and β-actin was used as a loading control, n = 3 ( c ). ( d ) Representative images of HOK, CAL27, and HSC-3 cells stained with ABCB5 (red), scale bar: 100 μm. ( e ) The ABCB5-positive areas, n = 5 fields. Data represent the mean ± SD; one-way ANOVA was used ( a,c,e )

Article Snippet: After blocking with 5% skimmed milk, the primary ABCB5 antibody (Santa Cruz Biotechnology, Inc., sc-515910, Shanghai, China) was incubated with a dilution of 1:500 for an entire night at 4°C.

Techniques: Quantitative RT-PCR, Western Blot, Control, Staining

Journal: Oncology Research

Article Title: Identifying ATP-Binding Cassette Member B5 as a New Biomarker for Oral Squamous Cell Carcinoma

doi: 10.32604/or.2025.064276

Figure Lengend Snippet: ABCB5 knockdown inhibits the migration and invasion of 2D OSCC cells: ( a–f ) ABCB5 was knocked down in the CAL27 cells using siABCB5, and the migration and invasion ability of the CAL27 cells were evaluated. ( a ) ABCB5 mRNA levels in CAL27 cells transfected with siABCB5 were evaluated using qRT-PCR, n = 3 qRT-PCR runs. ( b ) Representative images of CAL27 cells stained with ABCB5 (red), scale bar: 100 μm. ( c ) The ABCB5-positive areas, n = 5 fields. ( d–f ) The number of migrated ( d(upper),e ) and invaded ( d(lower),f ) CAL27 cells were imaged, scale bars: 100 μm ( d ) and counted, n = 5 fields ( e,f ). Data represent the mean ± SD; one-way ANOVA was used ( a,c,e,f )

Article Snippet: After blocking with 5% skimmed milk, the primary ABCB5 antibody (Santa Cruz Biotechnology, Inc., sc-515910, Shanghai, China) was incubated with a dilution of 1:500 for an entire night at 4°C.

Techniques: Knockdown, Migration, Transfection, Quantitative RT-PCR, Staining

Journal: Oncology Research

Article Title: Identifying ATP-Binding Cassette Member B5 as a New Biomarker for Oral Squamous Cell Carcinoma

doi: 10.32604/or.2025.064276

Figure Lengend Snippet: ABCB5 knockdown inhibits the migration and invasion of 3D OSCC spheroid: ( a–d ) The migration and invasion ability of 3D OSCC spheroid were evaluated in the CAL27 cells after transfected with siABCB5. ( a,b ) Representative images of migrated ( a ) and invaded ( b ) 3D OSCC spheroid, scale bars: 100 μm ( a,b ), and the area of cells migrated or invaded were calculated, n = 5 fields ( c,d ). Data represent the mean ± SD; one-way ANOVA was used ( c,d )

Article Snippet: After blocking with 5% skimmed milk, the primary ABCB5 antibody (Santa Cruz Biotechnology, Inc., sc-515910, Shanghai, China) was incubated with a dilution of 1:500 for an entire night at 4°C.

Techniques: Knockdown, Migration, Transfection

Journal: Oncology Research

Article Title: Identifying ATP-Binding Cassette Member B5 as a New Biomarker for Oral Squamous Cell Carcinoma

doi: 10.32604/or.2025.064276

Figure Lengend Snippet: ABCB5 Knockdown suppresses TGF-β-induced EMT: ( a ) The correlations between ABCB5 and epithelial marker E-cadherin and mesenchymal markers (N-cadherin and Vimentin) were observed by qRT-PCR in CAL27 (left) and HSC-3 (right) cells, which were treated with TGF-β in different concentrations, n = 3 qRT-PCR runs. ( b,c ) The mRNA expressions of epithelial marker E-cadherin ( b ) and mesenchymal markers (N-cadherin and Vimentin) ( c ) in CAL27 cells were analyzed using qRT-PCR, n = 3 qRT-PCR runs. ( d,e ) Representative images of CAL27 cells with or without TGF-β treatment were stained with E-cadherin, scale bar: 100 μm ( d ), and the positive areas of E-cadherin were calculated, n = 5 fields ( e ). ( f,g ) Representative images of CAL27 cells with or without TGF-β treatment were stained with Vimentin, scale bar: 100 μm ( f ), and the Vimentin-positive areas were quantified, n = 5 fields ( g ). Data represent the mean ± SD; one-way ANOVA was used ( b,c,e,g ); two-way ANOVA was used ( a ).

Article Snippet: After blocking with 5% skimmed milk, the primary ABCB5 antibody (Santa Cruz Biotechnology, Inc., sc-515910, Shanghai, China) was incubated with a dilution of 1:500 for an entire night at 4°C.

Techniques: Knockdown, Marker, Quantitative RT-PCR, Staining

Journal: Oncology Research

Article Title: Identifying ATP-Binding Cassette Member B5 as a New Biomarker for Oral Squamous Cell Carcinoma

doi: 10.32604/or.2025.064276

Figure Lengend Snippet: ABCB5 was highly expressed in the 4NQO-induced OSCC model in vivo: ( a–g ) The 4NQO carcinogen mouse model was established, and the expressions of ABCB5 and EMT markers were detected. ( a ) Schematic explains the experimental design of the 4NQO carcinogen mouse model. ( b ) Representative images of H&E staining, scale bar: 100 μm. ( c ) Representative image of tongue tissues stained with ABCB5, scale bar: 100 μm. ( d ) Quantification of ABCB5-positive areas, n = 5 fields. ( e ) mRNA expression of ABCB5 detected by qRT-PCR, n = 3 qRT-PCR runs. ( f,g ) mRNA levels of epithelial marker E-cadherin ( f ) and mesenchymal markers (N-cadherin and Vimentin) ( g ) were evaluated by qRT-PCR, n = 3 qRT-PCR runs, n = 5–8 mice per group. Data represent the mean ± SD; one-way ANOVA was used ( d,e,f,g )

Article Snippet: After blocking with 5% skimmed milk, the primary ABCB5 antibody (Santa Cruz Biotechnology, Inc., sc-515910, Shanghai, China) was incubated with a dilution of 1:500 for an entire night at 4°C.

Techniques: In Vivo, Staining, Expressing, Quantitative RT-PCR, Marker

Journal: Revista Brasileira de Ortopedia

Article Title: Osteopontin, WNT3A, and ABCB5 Biomarkers Expression in Osteosarcoma Patients

doi: 10.1055/s-0044-1788671

Figure Lengend Snippet: Description of primary antibodies, their respective manufacturers, and dilutions

Article Snippet: ABCB5 , Anti-ABCB5, clone 5H3C6 , GeneTex , 1:200.

Techniques:

Journal: Revista Brasileira de Ortopedia

Article Title: Osteopontin, WNT3A, and ABCB5 Biomarkers Expression in Osteosarcoma Patients

doi: 10.1055/s-0044-1788671

Figure Lengend Snippet: Descrição dos anticorpos primários com seus respectivos fabricantes e diluições

Article Snippet: ABCB5 , Anti-ABCB5, clone 5H3C6 , GeneTex , 1:200.

Techniques: