p her2  (Cell Signaling Technology Inc)


Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc p her2
    Assessment of <t>HER2</t> expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )
    P Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p her2/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p her2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines"

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    Journal: Journal of Nanobiotechnology

    doi: 10.1186/s12951-024-02600-7

    Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )
    Figure Legend Snippet: Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )

    Techniques Used: Expressing, Western Blot, Derivative Assay, Comparison, Staining, Flow Cytometry, Fluorescence

    Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis
    Figure Legend Snippet: Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis

    Techniques Used: Biomarker Assay, Membrane

    Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively
    Figure Legend Snippet: Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively

    Techniques Used: Derivative Assay, Expressing

    HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines
    Figure Legend Snippet: HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines

    Techniques Used: Western Blot, Clone Assay, Isolation, Selection, Stable Transfection, Transfection, shRNA, Flow Cytometry, Fluorescence, Biomarker Assay, Membrane

    Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown
    Figure Legend Snippet: Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown

    Techniques Used: shRNA, Expressing

    her2  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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  • 86

    Structured Review

    Cell Signaling Technology Inc her2
    Assessment of <t>HER2</t> expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )
    Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/her2/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    her2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines"

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    Journal: Journal of Nanobiotechnology

    doi: 10.1186/s12951-024-02600-7

    Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )
    Figure Legend Snippet: Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )

    Techniques Used: Expressing, Western Blot, Derivative Assay, Comparison, Staining, Flow Cytometry, Fluorescence

    Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis
    Figure Legend Snippet: Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis

    Techniques Used: Biomarker Assay, Membrane

    Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively
    Figure Legend Snippet: Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively

    Techniques Used: Derivative Assay, Expressing

    HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines
    Figure Legend Snippet: HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines

    Techniques Used: Western Blot, Clone Assay, Isolation, Selection, Stable Transfection, Transfection, shRNA, Flow Cytometry, Fluorescence, Biomarker Assay, Membrane

    Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown
    Figure Legend Snippet: Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown

    Techniques Used: shRNA, Expressing

    soluble her2  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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  • 86

    Structured Review

    Cell Signaling Technology Inc soluble her2
    Assessment of <t>HER2</t> expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )
    Soluble Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/soluble her2/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    soluble her2 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines"

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    Journal: Journal of Nanobiotechnology

    doi: 10.1186/s12951-024-02600-7

    Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )
    Figure Legend Snippet: Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )

    Techniques Used: Expressing, Western Blot, Derivative Assay, Comparison, Staining, Flow Cytometry, Fluorescence

    Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis
    Figure Legend Snippet: Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis

    Techniques Used: Biomarker Assay, Membrane

    Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively
    Figure Legend Snippet: Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively

    Techniques Used: Derivative Assay, Expressing

    HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines
    Figure Legend Snippet: HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines

    Techniques Used: Western Blot, Clone Assay, Isolation, Selection, Stable Transfection, Transfection, shRNA, Flow Cytometry, Fluorescence, Biomarker Assay, Membrane

    Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown
    Figure Legend Snippet: Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown

    Techniques Used: shRNA, Expressing

    her2  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc her2
    Protein expression profiles of MCF7 LTLT cells versus normal MCF7, T47D, and MCF7aro cells. The cells were assayed by immunoblot with antibodies against ( A ) ERα; ( B ) <t>HER2;</t> ( C ) GR; ( D ) AR; ( E ) PR. β-actin was used as an internal loading control. For AR and HER2, samples were run on a WES (ProteinSimple). For ERα, GR, and PR, one of two representative experiments were shown
    Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Lasofoxifene as a potential treatment for aromatase inhibitor-resistant ER-positive breast cancer"

    Article Title: Lasofoxifene as a potential treatment for aromatase inhibitor-resistant ER-positive breast cancer

    Journal: Breast Cancer Research : BCR

    doi: 10.1186/s13058-024-01843-4

    Protein expression profiles of MCF7 LTLT cells versus normal MCF7, T47D, and MCF7aro cells. The cells were assayed by immunoblot with antibodies against ( A ) ERα; ( B ) HER2; ( C ) GR; ( D ) AR; ( E ) PR. β-actin was used as an internal loading control. For AR and HER2, samples were run on a WES (ProteinSimple). For ERα, GR, and PR, one of two representative experiments were shown
    Figure Legend Snippet: Protein expression profiles of MCF7 LTLT cells versus normal MCF7, T47D, and MCF7aro cells. The cells were assayed by immunoblot with antibodies against ( A ) ERα; ( B ) HER2; ( C ) GR; ( D ) AR; ( E ) PR. β-actin was used as an internal loading control. For AR and HER2, samples were run on a WES (ProteinSimple). For ERα, GR, and PR, one of two representative experiments were shown

    Techniques Used: Expressing, Western Blot

    anti her2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti her2
    Immunoblotting evaluating the activation of STAT3 and <t>HER2</t> in MCF-7 ( A ), STAT3 and EGFR in H358 ( B ), STAT3 and SRC in LNCAP ( C ), and STAT3 and JACK2 in HepG2 ( D ). Total protein extracts were subjected to immunoblot analysis. Immunoblot evidenced that STAT3 phosphorylation (pY705-STAT3), HER2 phosphorylation (pY1248-HER2), JACK2 phosphorylation (pY1007/1008-JAK2), and Src phosphorylation (pY416-Src) increased upon treatment with 10 µM of β-HCH. The increase in STAT3 phosphorylation was upheld even with the combined treatment β-HCH+ TKIs, whereas a decrease in the band intensity occurred in the sample with triple treatment (β-HCH+ TKI+ S3I-201), as evident in the relative densitometry. β-actin was used for housekeeping. Phosphorylation levels referred to the amount of total STAT3, HER2, EGFR, Src, or JAK2 present in each sample and were compared with the control. These images are representative of three independent experiments with similar results. Statistical analysis was performed with the GraphPad Prisma software using ANOVA followed by Tukey’s post hoc test. Statistically significant differences referring to the CTRL or TKIs are marked with asterisks (ns: not statistically significant, ** p < 0.01; *** p < 0.001; **** p < 0.0001).
    Anti Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "STAT3 Pathways Contribute to β-HCH Interference with Anticancer Tyrosine Kinase Inhibitors"

    Article Title: STAT3 Pathways Contribute to β-HCH Interference with Anticancer Tyrosine Kinase Inhibitors

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms25116181

    Immunoblotting evaluating the activation of STAT3 and HER2 in MCF-7 ( A ), STAT3 and EGFR in H358 ( B ), STAT3 and SRC in LNCAP ( C ), and STAT3 and JACK2 in HepG2 ( D ). Total protein extracts were subjected to immunoblot analysis. Immunoblot evidenced that STAT3 phosphorylation (pY705-STAT3), HER2 phosphorylation (pY1248-HER2), JACK2 phosphorylation (pY1007/1008-JAK2), and Src phosphorylation (pY416-Src) increased upon treatment with 10 µM of β-HCH. The increase in STAT3 phosphorylation was upheld even with the combined treatment β-HCH+ TKIs, whereas a decrease in the band intensity occurred in the sample with triple treatment (β-HCH+ TKI+ S3I-201), as evident in the relative densitometry. β-actin was used for housekeeping. Phosphorylation levels referred to the amount of total STAT3, HER2, EGFR, Src, or JAK2 present in each sample and were compared with the control. These images are representative of three independent experiments with similar results. Statistical analysis was performed with the GraphPad Prisma software using ANOVA followed by Tukey’s post hoc test. Statistically significant differences referring to the CTRL or TKIs are marked with asterisks (ns: not statistically significant, ** p < 0.01; *** p < 0.001; **** p < 0.0001).
    Figure Legend Snippet: Immunoblotting evaluating the activation of STAT3 and HER2 in MCF-7 ( A ), STAT3 and EGFR in H358 ( B ), STAT3 and SRC in LNCAP ( C ), and STAT3 and JACK2 in HepG2 ( D ). Total protein extracts were subjected to immunoblot analysis. Immunoblot evidenced that STAT3 phosphorylation (pY705-STAT3), HER2 phosphorylation (pY1248-HER2), JACK2 phosphorylation (pY1007/1008-JAK2), and Src phosphorylation (pY416-Src) increased upon treatment with 10 µM of β-HCH. The increase in STAT3 phosphorylation was upheld even with the combined treatment β-HCH+ TKIs, whereas a decrease in the band intensity occurred in the sample with triple treatment (β-HCH+ TKI+ S3I-201), as evident in the relative densitometry. β-actin was used for housekeeping. Phosphorylation levels referred to the amount of total STAT3, HER2, EGFR, Src, or JAK2 present in each sample and were compared with the control. These images are representative of three independent experiments with similar results. Statistical analysis was performed with the GraphPad Prisma software using ANOVA followed by Tukey’s post hoc test. Statistically significant differences referring to the CTRL or TKIs are marked with asterisks (ns: not statistically significant, ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Techniques Used: Western Blot, Activation Assay, Software

    phosphor her2 tyr1221 1222  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phosphor her2 tyr1221 1222
    Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)
    Phosphor Her2 Tyr1221 1222, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Synergistic Effects of Neratinib in Combination With Palbociclib or Miransertib in Brain Cancer Cells"

    Article Title: Synergistic Effects of Neratinib in Combination With Palbociclib or Miransertib in Brain Cancer Cells

    Journal: World Journal of Oncology

    doi: 10.14740/wjon1873

    Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)
    Figure Legend Snippet: Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)

    Techniques Used: Expressing, Cytometry, Fluorescence

    IC 50 Values of Various Agents on HBCCLs as Assessed by SRB Colorimetric Assay: (A) HER-Family Targeting TKIs and Other Downstream Signaling Molecules and (B) Other TKIs and Chemotherapeutic Agents
    Figure Legend Snippet: IC 50 Values of Various Agents on HBCCLs as Assessed by SRB Colorimetric Assay: (A) HER-Family Targeting TKIs and Other Downstream Signaling Molecules and (B) Other TKIs and Chemotherapeutic Agents

    Techniques Used: Colorimetric Assay, Polymer

    Linear Regression Analysis of the Expression of Various Receptors Against the Sensitivity of Human Brain Cancer Cell Lines to Treatment With Various TKIs, CDK Inhibitors, STAT3 Inhibitor and Cytotoxic Agents
    Figure Legend Snippet: Linear Regression Analysis of the Expression of Various Receptors Against the Sensitivity of Human Brain Cancer Cell Lines to Treatment With Various TKIs, CDK Inhibitors, STAT3 Inhibitor and Cytotoxic Agents

    Techniques Used: Expressing, Significance Assay

    her2  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc her2
    Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)
    Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Synergistic Effects of Neratinib in Combination With Palbociclib or Miransertib in Brain Cancer Cells"

    Article Title: Synergistic Effects of Neratinib in Combination With Palbociclib or Miransertib in Brain Cancer Cells

    Journal: World Journal of Oncology

    doi: 10.14740/wjon1873

    Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)
    Figure Legend Snippet: Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)

    Techniques Used: Expressing, Cytometry, Fluorescence

    IC 50 Values of Various Agents on HBCCLs as Assessed by SRB Colorimetric Assay: (A) HER-Family Targeting TKIs and Other Downstream Signaling Molecules and (B) Other TKIs and Chemotherapeutic Agents
    Figure Legend Snippet: IC 50 Values of Various Agents on HBCCLs as Assessed by SRB Colorimetric Assay: (A) HER-Family Targeting TKIs and Other Downstream Signaling Molecules and (B) Other TKIs and Chemotherapeutic Agents

    Techniques Used: Colorimetric Assay, Polymer

    Linear Regression Analysis of the Expression of Various Receptors Against the Sensitivity of Human Brain Cancer Cell Lines to Treatment With Various TKIs, CDK Inhibitors, STAT3 Inhibitor and Cytotoxic Agents
    Figure Legend Snippet: Linear Regression Analysis of the Expression of Various Receptors Against the Sensitivity of Human Brain Cancer Cell Lines to Treatment With Various TKIs, CDK Inhibitors, STAT3 Inhibitor and Cytotoxic Agents

    Techniques Used: Expressing, Significance Assay

    her2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc her2
    Clonogenic cell survival and immunoblot assays of <t>HER2</t> - and MET -amplified OE33 cells. ( a ) Cells were treated with trastuzumab and capmatinib, alone and in combination, for 14 d, and clonogenic assays were performed. ( b ) Immunoblotting analysis using antibodies specific for the proteins of lysates treated with trastuzumab and savolitinib alone or in combination for 24 h before harvest. Combined treatment with savolitinib inhibited the activation of extracellular signal-regulated kinase (ERK)-1/2 and protein kinase B (AKT) via MET dephosphorylation in OE33 cells.
    Her2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Characterization of MET Alterations in 37 Gastroesophageal Cancer Cell Lines for MET-Targeted Therapy"

    Article Title: Characterization of MET Alterations in 37 Gastroesophageal Cancer Cell Lines for MET-Targeted Therapy

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms25115975

    Clonogenic cell survival and immunoblot assays of HER2 - and MET -amplified OE33 cells. ( a ) Cells were treated with trastuzumab and capmatinib, alone and in combination, for 14 d, and clonogenic assays were performed. ( b ) Immunoblotting analysis using antibodies specific for the proteins of lysates treated with trastuzumab and savolitinib alone or in combination for 24 h before harvest. Combined treatment with savolitinib inhibited the activation of extracellular signal-regulated kinase (ERK)-1/2 and protein kinase B (AKT) via MET dephosphorylation in OE33 cells.
    Figure Legend Snippet: Clonogenic cell survival and immunoblot assays of HER2 - and MET -amplified OE33 cells. ( a ) Cells were treated with trastuzumab and capmatinib, alone and in combination, for 14 d, and clonogenic assays were performed. ( b ) Immunoblotting analysis using antibodies specific for the proteins of lysates treated with trastuzumab and savolitinib alone or in combination for 24 h before harvest. Combined treatment with savolitinib inhibited the activation of extracellular signal-regulated kinase (ERK)-1/2 and protein kinase B (AKT) via MET dephosphorylation in OE33 cells.

    Techniques Used: Western Blot, Amplification, Activation Assay, De-Phosphorylation Assay

    p-her2-tyr 122  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p-her2-tyr 122
    P Her2 Tyr 122, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    her2 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc her2 antibody
    Her2 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/her2 antibody/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc p her2
    Assessment of <t>HER2</t> expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )
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    Assessment of <t>HER2</t> expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )
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    Cell Signaling Technology Inc soluble her2
    Assessment of <t>HER2</t> expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )
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    Cell Signaling Technology Inc anti her2
    Immunoblotting evaluating the activation of STAT3 and <t>HER2</t> in MCF-7 ( A ), STAT3 and EGFR in H358 ( B ), STAT3 and SRC in LNCAP ( C ), and STAT3 and JACK2 in HepG2 ( D ). Total protein extracts were subjected to immunoblot analysis. Immunoblot evidenced that STAT3 phosphorylation (pY705-STAT3), HER2 phosphorylation (pY1248-HER2), JACK2 phosphorylation (pY1007/1008-JAK2), and Src phosphorylation (pY416-Src) increased upon treatment with 10 µM of β-HCH. The increase in STAT3 phosphorylation was upheld even with the combined treatment β-HCH+ TKIs, whereas a decrease in the band intensity occurred in the sample with triple treatment (β-HCH+ TKI+ S3I-201), as evident in the relative densitometry. β-actin was used for housekeeping. Phosphorylation levels referred to the amount of total STAT3, HER2, EGFR, Src, or JAK2 present in each sample and were compared with the control. These images are representative of three independent experiments with similar results. Statistical analysis was performed with the GraphPad Prisma software using ANOVA followed by Tukey’s post hoc test. Statistically significant differences referring to the CTRL or TKIs are marked with asterisks (ns: not statistically significant, ** p < 0.01; *** p < 0.001; **** p < 0.0001).
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    Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)
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    Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)
    P Her2 Tyr 122, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)
    Her2 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Expressing, Western Blot, Derivative Assay, Comparison, Staining, Flow Cytometry, Fluorescence

    Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Biomarker Assay, Membrane

    Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Derivative Assay, Expressing

    HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Western Blot, Clone Assay, Isolation, Selection, Stable Transfection, Transfection, shRNA, Flow Cytometry, Fluorescence, Biomarker Assay, Membrane

    Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: shRNA, Expressing

    Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Expressing, Western Blot, Derivative Assay, Comparison, Staining, Flow Cytometry, Fluorescence

    Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Biomarker Assay, Membrane

    Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Derivative Assay, Expressing

    HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: Western Blot, Clone Assay, Isolation, Selection, Stable Transfection, Transfection, shRNA, Flow Cytometry, Fluorescence, Biomarker Assay, Membrane

    Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown

    Article Snippet: Antibodies to p-HER2 (Y1248 # 2247), HER2 (#3250), p-AKT (S473) (#9271), AKT (#9272) were from Cell Signaling Technology (Beverly, MA USA); α-Tubulin (T5168) was from Sigma-Aldrich Co. (Merck KGaA, St. Louis, MO, USA).

    Techniques: shRNA, Expressing

    Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Assessment of HER2 expression in a series of breast cell lines. ( A ) List of the cell lines employed in this study ( B ) Western blot analysis of HER2 expression on extracts from MDA-MB-231, MDA-MB-468, MCF7, BT474 and SKBR3 cells derived from different BCs; MCF10A cells, derived from non-tumorigenic breast epithelium, were used for comparison. α-Tubulin was used to normalise the amounts of proteins loaded in each lane. The histogram reports HER2 total expression as Relative Units (R.U.) compared to MCF10A cells. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( C ) Western blot analysis of the soluble form of HER2 (sHER2) present in the concentrated media from the same cells as in B). The histogram shows the relative quantification, expressed as Relative Units (R.U.), after normalization to protein loading through Ponceau S staining of the filter. Statistical significance is considered when **** p < 0.0001 ( ANOVA ). ( D ) Flow cytometry analysis of HER2 cell surface exposure in the same cell lines as in A). The percentages of cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The plots also report the positions of the isotypes as black lines. The histogram shows the relative quantification, expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( ANOVA )

    Article Snippet: Soluble HER2 was detected with an antibody against the N-terminal part of the receptor (#4290 Cell Signaling Technology, Inc., Beverly, MA USA).

    Techniques: Expressing, Western Blot, Derivative Assay, Comparison, Staining, Flow Cytometry, Fluorescence

    Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Brightfield imaged and SERS intensity maps of a selected cell for the Raman peaks at 1080 and 1580 cm − 1 for the cell lines: ( A ) MCF10A ( B ) MDA-MB-468 ( C ) SKBR3. Scale bar = 10 μm. ( D ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines based on the SERS analysis

    Article Snippet: Soluble HER2 was detected with an antibody against the N-terminal part of the receptor (#4290 Cell Signaling Technology, Inc., Beverly, MA USA).

    Techniques: Biomarker Assay, Membrane

    Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Raman classification of breast derived cell lines expressing different HER2 levels. ( A ) Median + IQR spectra of SKBR3 (lime), MCF10A (green) and MDA-MB-468 (blue), with PCA loading expressing high variance, black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) 3D score plot of PC2-4 scores demonstrating the classification achieved for SKBR3, MCF10A and MDA-MB-468 cells using 3 components. ( C ) Box-plot of the distribution for each cell line of PC4 scores. The asterisks represent Wilcoxon nonparametric test results with p < 0.0001 when ****. ( D - E ) LDA supervised classification with the separation achieved using the latent variables 1 and 2, and the misclassification rate, respectively

    Article Snippet: Soluble HER2 was detected with an antibody against the N-terminal part of the receptor (#4290 Cell Signaling Technology, Inc., Beverly, MA USA).

    Techniques: Derivative Assay, Expressing

    HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: HER2 assessment in scrambled- and HER2-silenced SKBR3 cells. ( A ) Western blot analysis of HER2 on extracts from single clones isolated upon selection of SKBR3 stably transfected with a scrambled control (SCR) or with an shRNA directed against HER2 (HER2 shRNA Clones 1–3). α-Tubulin was used as loading control for each sample. ( B ) Flow cytometry analysis of HER2 cell surface exposure in the scrambled transfected SKBR3 (SCR) or in clone 3 (Clone 3). The cell counts are reported on the y axis, while the Fluorescence Intensity on the x axis. The histogram shows the relative quantification expressed as Mean Fluorescence Intensity (MFI). Statistical significance is considered when **** p < 0.0001 ( t -test). ( C ) Western blot analysis of HER2, AKT and the phosphorylated form levels in extracts from SKBR3 transfected with a scrambled control and from the HER2-silenced clone 3. The first histogram illustrates the quantification of HER2 relative to α-Tubulin and the second the AKT phosphorylation level to total AKT compared to the SCR control. Statistical significance is considered when *** p < 0.001 and **** p < 0.0001 ( t -test). ( D ) Brightfield images and SERS intensity maps of a selected cell for the peaks at 1080 and 1580 cm − 1 for scrambled- and HER2-silenced SKBR3 cells. Scale bar = 10 μm. ( E ) Relative quantification of HER2 biomarker on cell membrane of the analysed cell lines

    Article Snippet: Soluble HER2 was detected with an antibody against the N-terminal part of the receptor (#4290 Cell Signaling Technology, Inc., Beverly, MA USA).

    Techniques: Western Blot, Clone Assay, Isolation, Selection, Stable Transfection, Transfection, shRNA, Flow Cytometry, Fluorescence, Biomarker Assay, Membrane

    Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown

    Journal: Journal of Nanobiotechnology

    Article Title: Combined SERS-Raman screening of HER2-overexpressing or silenced breast cancer cell lines

    doi: 10.1186/s12951-024-02600-7

    Figure Lengend Snippet: Raman spectra of the scrambled- and HER2-silenced SKBR3 cells (clone 3). ( A ): Median IQR spectra of scrambled (lime) and HER2-shRNA SKBR3 (red) with PCA loadings expressing high variance, as black lines (y is offset for clarity). Vertical dash lines are single peaks of interest; cyan vertical bands are relevant biological bands in cell classification. ( B ) The box plot of distribution of PCA scores for PC4 for the scrambled SKBR3 and silenced counterpart is shown, with a Wilcoxon statistic. ( C ) The score plot of PC1-PC4, evidencing the best spatial separation among scrambled and silenced SKBR3, is shown

    Article Snippet: Soluble HER2 was detected with an antibody against the N-terminal part of the receptor (#4290 Cell Signaling Technology, Inc., Beverly, MA USA).

    Techniques: shRNA, Expressing

    Immunoblotting evaluating the activation of STAT3 and HER2 in MCF-7 ( A ), STAT3 and EGFR in H358 ( B ), STAT3 and SRC in LNCAP ( C ), and STAT3 and JACK2 in HepG2 ( D ). Total protein extracts were subjected to immunoblot analysis. Immunoblot evidenced that STAT3 phosphorylation (pY705-STAT3), HER2 phosphorylation (pY1248-HER2), JACK2 phosphorylation (pY1007/1008-JAK2), and Src phosphorylation (pY416-Src) increased upon treatment with 10 µM of β-HCH. The increase in STAT3 phosphorylation was upheld even with the combined treatment β-HCH+ TKIs, whereas a decrease in the band intensity occurred in the sample with triple treatment (β-HCH+ TKI+ S3I-201), as evident in the relative densitometry. β-actin was used for housekeeping. Phosphorylation levels referred to the amount of total STAT3, HER2, EGFR, Src, or JAK2 present in each sample and were compared with the control. These images are representative of three independent experiments with similar results. Statistical analysis was performed with the GraphPad Prisma software using ANOVA followed by Tukey’s post hoc test. Statistically significant differences referring to the CTRL or TKIs are marked with asterisks (ns: not statistically significant, ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Journal: International Journal of Molecular Sciences

    Article Title: STAT3 Pathways Contribute to β-HCH Interference with Anticancer Tyrosine Kinase Inhibitors

    doi: 10.3390/ijms25116181

    Figure Lengend Snippet: Immunoblotting evaluating the activation of STAT3 and HER2 in MCF-7 ( A ), STAT3 and EGFR in H358 ( B ), STAT3 and SRC in LNCAP ( C ), and STAT3 and JACK2 in HepG2 ( D ). Total protein extracts were subjected to immunoblot analysis. Immunoblot evidenced that STAT3 phosphorylation (pY705-STAT3), HER2 phosphorylation (pY1248-HER2), JACK2 phosphorylation (pY1007/1008-JAK2), and Src phosphorylation (pY416-Src) increased upon treatment with 10 µM of β-HCH. The increase in STAT3 phosphorylation was upheld even with the combined treatment β-HCH+ TKIs, whereas a decrease in the band intensity occurred in the sample with triple treatment (β-HCH+ TKI+ S3I-201), as evident in the relative densitometry. β-actin was used for housekeeping. Phosphorylation levels referred to the amount of total STAT3, HER2, EGFR, Src, or JAK2 present in each sample and were compared with the control. These images are representative of three independent experiments with similar results. Statistical analysis was performed with the GraphPad Prisma software using ANOVA followed by Tukey’s post hoc test. Statistically significant differences referring to the CTRL or TKIs are marked with asterisks (ns: not statistically significant, ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Article Snippet: The antibodies used in the Western blotting analysis are listed as follows: anti-STAT3 (Cell Signaling, Pero, Italy, Cat. No. 9132), anti-pY705-STAT3 (Cell Signaling, Pero, Italy, Cat. No. 9145), anti-p-SRC (Cell Signaling, Pero, Italy, Cat. No. 6943), anti-Src (Cell Signaling, Pero, Italy, Cat. No. 2108), anti-β-actin (Sigma-Aldrich, Milan, Italy, Cat. No. A1978 clone AC-15), anti-pY1007/1008-Jack2 (Cell Signaling, Pero, Italy, Cat. No. 3776), anti-Jack2 (Cell Signaling, Pero, Italy, Cat. No. 3230), anti-pHER2 (Cell Signaling, Pero, Italy, Cat. No. 2247S), and anti-HER2 (Cell Signaling, Pero, Italy, Cat. No. 2242).

    Techniques: Western Blot, Activation Assay, Software

    Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)

    Journal: World Journal of Oncology

    Article Title: Synergistic Effects of Neratinib in Combination With Palbociclib or Miransertib in Brain Cancer Cells

    doi: 10.14740/wjon1873

    Figure Lengend Snippet: Surface Expression of Various Growth Factor Receptors in Human Brain Cancer Cell Line (Determined by Flow Cytometry)

    Article Snippet: Other antibodies for the Western blot analysis, such as the rabbit anti-phospho-EGFR (Tyr1068), HER2 (2242s), phosphor-HER2 (Tyr1221/1222) (2243), phosphor-HER3 (Tyr1289) (4791), phosphor-HER4 (Tyr1284)/EGFR (Tyr1173) (4757), phosphor-MAPK (Tyr202/Tyr204) (4370), phospho-Akt (S473) (4060), phospho-STAT3 (Y705) (9145), phospho-SRC (Y416) (Tyr416) (6942) and β-actin (4970), were all obtained from Cell Signaling Technology Inc. (Hitchin, UK).

    Techniques: Expressing, Cytometry, Fluorescence

    IC 50 Values of Various Agents on HBCCLs as Assessed by SRB Colorimetric Assay: (A) HER-Family Targeting TKIs and Other Downstream Signaling Molecules and (B) Other TKIs and Chemotherapeutic Agents

    Journal: World Journal of Oncology

    Article Title: Synergistic Effects of Neratinib in Combination With Palbociclib or Miransertib in Brain Cancer Cells

    doi: 10.14740/wjon1873

    Figure Lengend Snippet: IC 50 Values of Various Agents on HBCCLs as Assessed by SRB Colorimetric Assay: (A) HER-Family Targeting TKIs and Other Downstream Signaling Molecules and (B) Other TKIs and Chemotherapeutic Agents

    Article Snippet: Other antibodies for the Western blot analysis, such as the rabbit anti-phospho-EGFR (Tyr1068), HER2 (2242s), phosphor-HER2 (Tyr1221/1222) (2243), phosphor-HER3 (Tyr1289) (4791), phosphor-HER4 (Tyr1284)/EGFR (Tyr1173) (4757), phosphor-MAPK (Tyr202/Tyr204) (4370), phospho-Akt (S473) (4060), phospho-STAT3 (Y705) (9145), phospho-SRC (Y416) (Tyr416) (6942) and β-actin (4970), were all obtained from Cell Signaling Technology Inc. (Hitchin, UK).

    Techniques: Colorimetric Assay, Polymer

    Linear Regression Analysis of the Expression of Various Receptors Against the Sensitivity of Human Brain Cancer Cell Lines to Treatment With Various TKIs, CDK Inhibitors, STAT3 Inhibitor and Cytotoxic Agents

    Journal: World Journal of Oncology

    Article Title: Synergistic Effects of Neratinib in Combination With Palbociclib or Miransertib in Brain Cancer Cells

    doi: 10.14740/wjon1873

    Figure Lengend Snippet: Linear Regression Analysis of the Expression of Various Receptors Against the Sensitivity of Human Brain Cancer Cell Lines to Treatment With Various TKIs, CDK Inhibitors, STAT3 Inhibitor and Cytotoxic Agents

    Article Snippet: Other antibodies for the Western blot analysis, such as the rabbit anti-phospho-EGFR (Tyr1068), HER2 (2242s), phosphor-HER2 (Tyr1221/1222) (2243), phosphor-HER3 (Tyr1289) (4791), phosphor-HER4 (Tyr1284)/EGFR (Tyr1173) (4757), phosphor-MAPK (Tyr202/Tyr204) (4370), phospho-Akt (S473) (4060), phospho-STAT3 (Y705) (9145), phospho-SRC (Y416) (Tyr416) (6942) and β-actin (4970), were all obtained from Cell Signaling Technology Inc. (Hitchin, UK).

    Techniques: Expressing, Significance Assay