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: Advanced healthcare materials

Article Title: Hydrogel microtumor arrays to evaluate nanotherapeutics

doi: 10.1002/adhm.202201696

Figure Lengend Snippet: Mechanical cues prime a population of cells with increase stemness which increase uptake of Cy5-PFC3 (A) Representative image of fluorescence intensity quantitation in B16F0 cells (Image J). Cells outlined in red considered ‘perimeter’ and yellow considered ‘centre’. Heatmap of nanoparticle intensity after five days culture (n=10). (B) Scatter plot of fluorescence intensity of B16F0 cells localised at the perimeter and the centre of a pattern cultured for five days after 1 h treatment with Cy5-PFC3. Significant difference in uptake between cells in perimeter and centre (p = 0.0002, n = 12) and ABCB5 expression (p = 0.0079, n = 12). (C) Scatter plot of fluorescence intensity of B16F0 and B16F10 cells in spiral patterns cultured on 10 kPa polyacrylamide substrate. Error bars denote ± standard deviation. Significant difference in Cy5-PFC3 uptake between B16F0 1 day and 5 days (p = 0.0005, n = 15), B16F0 and B16F10 at 5 days (p < 0.0001, n = 15) and ABCB5 expression between B16F0 1 day and 5 day (p < 0.0001, n = 15), B16F0 and B16F10 at 5 days (p < 0.0174, n = 20)

Article Snippet: Gels were then incubated with the primary antibody (rabbit polyclonal anti-ABCB5, Novus Biologicals) diluted at 1:500 in 1% BSA in PBS for 1h at room temperature.

Techniques: Fluorescence, Quantitation Assay, Cell Culture, Expressing, Standard Deviation

Journal: Advanced healthcare materials

Article Title: Hydrogel microtumor arrays to evaluate nanotherapeutics

doi: 10.1002/adhm.202201696

Figure Lengend Snippet: Shape induced and innate cellular ABCB5 expression increases nanoparticle uptake in murine melanoma cells. (Scale bar = 100 μm) (A) Representative fluorescence images of a B16F0 one day culture, B16F0 five day culture, B16F10 one day culture, and B16F10 five day culture. Fluorescence channels present Cy5-PFC3 nanoparticle (Red), ABCB5 putative cancer stem cell marker (Alexa Fluor 555, Orange) and cancer cell nuclei (DAPI) on spiral patterns. (B) Quantification of mean Cy5-PFC3 and ABCB5 stem cell marker intensity in B16F0 (n = 10) and B16F10 (n = 10) cells proliferated under spiral geometry for one day or five days with standard deviation shown with error bars. ****: P ≤ 0.0001, ***: P ≤0.001

Article Snippet: Gels were then incubated with the primary antibody (rabbit polyclonal anti-ABCB5, Novus Biologicals) diluted at 1:500 in 1% BSA in PBS for 1h at room temperature.

Techniques: Expressing, Fluorescence, Marker, Standard Deviation

Journal: Cancer Drug Resistance

Article Title: The uniqueness of ABCB5 as a full transporter ABCB5FL and a half-transporter-like ABCB5β

doi: 10.20517/cdr.2024.56

Figure Lengend Snippet: Cancers in which ABCB5 expression has been identified outside of its role as a marker of CSC

Article Snippet: Immunohistochemistry using antibody NBP1-77687 from Novus Biological , [ ] .

Techniques: Expressing, Marker, Next-Generation Sequencing, Sequencing, Immunohistochemistry, DNA Sequencing

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