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Analysis of HIF-1α and CYP1B1 expression in tissue <t>microarray</t> (TMA) samples from breast cancer patients. (A) Schematic diagram of the tumor stages of each breast tissue core. (B) Hematoxylin and eosin (H&E) staining images provided by the manufacturer. Immunofluorescence images of tumor microarrays (TMA) stained with anti-hypoxia-inducible factor-1 alpha (HIF-1α) (C) and anti-cytochrome P450 1B1 (CYP1B1) (D) antibodies. Images were acquired at 20X magnification using a Leica Thunder microscope. Whole tissue slide images were mosaic merged via LAS-X software. (E) Representative images of immunofluorescence analysis of HIF-1α (green) and CYP1B1 (green) in two slides of ER-negative and positive breast cancer TMA. Immunofluorescence images were acquired using a Leica Thunder microscope. (F) Scatter plots showing fluorescence intensity correlation analysis of HIF-1α and CYP1B1 in 64 ER-positive TMAs. (G) Violin plots of fluorescence intensities of HIF-1α and CYP1B1. Samples were classified into low (immunohistochemical [IHC] score: 1+, 2+) and high (IHC score: 3+) groups according to the estrogen receptor (ER) expression status of the TMA specification sheet, and the correlation between HIF-1α and CYP1B1 expression was analyzed. (H) Analysis of fluorescence intensity of CYP1B1 against HIF1α expression in ER-positive TMA. The fluorescence intensity of CYP1B1 was analyzed by classifying the fluorescence intensity of HIF1-α based on the top and bottom 25% ER expression. *, P<0.0001 versus ER-positive tumor tissue with low HIF1-α fluorescence intensity by Mann-Whitney test.
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Analysis of HIF-1α and CYP1B1 expression in tissue <t>microarray</t> (TMA) samples from breast cancer patients. (A) Schematic diagram of the tumor stages of each breast tissue core. (B) Hematoxylin and eosin (H&E) staining images provided by the manufacturer. Immunofluorescence images of tumor microarrays (TMA) stained with anti-hypoxia-inducible factor-1 alpha (HIF-1α) (C) and anti-cytochrome P450 1B1 (CYP1B1) (D) antibodies. Images were acquired at 20X magnification using a Leica Thunder microscope. Whole tissue slide images were mosaic merged via LAS-X software. (E) Representative images of immunofluorescence analysis of HIF-1α (green) and CYP1B1 (green) in two slides of ER-negative and positive breast cancer TMA. Immunofluorescence images were acquired using a Leica Thunder microscope. (F) Scatter plots showing fluorescence intensity correlation analysis of HIF-1α and CYP1B1 in 64 ER-positive TMAs. (G) Violin plots of fluorescence intensities of HIF-1α and CYP1B1. Samples were classified into low (immunohistochemical [IHC] score: 1+, 2+) and high (IHC score: 3+) groups according to the estrogen receptor (ER) expression status of the TMA specification sheet, and the correlation between HIF-1α and CYP1B1 expression was analyzed. (H) Analysis of fluorescence intensity of CYP1B1 against HIF1α expression in ER-positive TMA. The fluorescence intensity of CYP1B1 was analyzed by classifying the fluorescence intensity of HIF1-α based on the top and bottom 25% ER expression. *, P<0.0001 versus ER-positive tumor tissue with low HIF1-α fluorescence intensity by Mann-Whitney test.
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Analysis of HIF-1α and CYP1B1 expression in tissue <t>microarray</t> (TMA) samples from breast cancer patients. (A) Schematic diagram of the tumor stages of each breast tissue core. (B) Hematoxylin and eosin (H&E) staining images provided by the manufacturer. Immunofluorescence images of tumor microarrays (TMA) stained with anti-hypoxia-inducible factor-1 alpha (HIF-1α) (C) and anti-cytochrome P450 1B1 (CYP1B1) (D) antibodies. Images were acquired at 20X magnification using a Leica Thunder microscope. Whole tissue slide images were mosaic merged via LAS-X software. (E) Representative images of immunofluorescence analysis of HIF-1α (green) and CYP1B1 (green) in two slides of ER-negative and positive breast cancer TMA. Immunofluorescence images were acquired using a Leica Thunder microscope. (F) Scatter plots showing fluorescence intensity correlation analysis of HIF-1α and CYP1B1 in 64 ER-positive TMAs. (G) Violin plots of fluorescence intensities of HIF-1α and CYP1B1. Samples were classified into low (immunohistochemical [IHC] score: 1+, 2+) and high (IHC score: 3+) groups according to the estrogen receptor (ER) expression status of the TMA specification sheet, and the correlation between HIF-1α and CYP1B1 expression was analyzed. (H) Analysis of fluorescence intensity of CYP1B1 against HIF1α expression in ER-positive TMA. The fluorescence intensity of CYP1B1 was analyzed by classifying the fluorescence intensity of HIF1-α based on the top and bottom 25% ER expression. *, P<0.0001 versus ER-positive tumor tissue with low HIF1-α fluorescence intensity by Mann-Whitney test.
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Analysis of HIF-1α and CYP1B1 expression in tissue <t>microarray</t> (TMA) samples from breast cancer patients. (A) Schematic diagram of the tumor stages of each breast tissue core. (B) Hematoxylin and eosin (H&E) staining images provided by the manufacturer. Immunofluorescence images of tumor microarrays (TMA) stained with anti-hypoxia-inducible factor-1 alpha (HIF-1α) (C) and anti-cytochrome P450 1B1 (CYP1B1) (D) antibodies. Images were acquired at 20X magnification using a Leica Thunder microscope. Whole tissue slide images were mosaic merged via LAS-X software. (E) Representative images of immunofluorescence analysis of HIF-1α (green) and CYP1B1 (green) in two slides of ER-negative and positive breast cancer TMA. Immunofluorescence images were acquired using a Leica Thunder microscope. (F) Scatter plots showing fluorescence intensity correlation analysis of HIF-1α and CYP1B1 in 64 ER-positive TMAs. (G) Violin plots of fluorescence intensities of HIF-1α and CYP1B1. Samples were classified into low (immunohistochemical [IHC] score: 1+, 2+) and high (IHC score: 3+) groups according to the estrogen receptor (ER) expression status of the TMA specification sheet, and the correlation between HIF-1α and CYP1B1 expression was analyzed. (H) Analysis of fluorescence intensity of CYP1B1 against HIF1α expression in ER-positive TMA. The fluorescence intensity of CYP1B1 was analyzed by classifying the fluorescence intensity of HIF1-α based on the top and bottom 25% ER expression. *, P<0.0001 versus ER-positive tumor tissue with low HIF1-α fluorescence intensity by Mann-Whitney test.
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Analysis of HIF-1α and CYP1B1 expression in tissue microarray (TMA) samples from breast cancer patients. (A) Schematic diagram of the tumor stages of each breast tissue core. (B) Hematoxylin and eosin (H&E) staining images provided by the manufacturer. Immunofluorescence images of tumor microarrays (TMA) stained with anti-hypoxia-inducible factor-1 alpha (HIF-1α) (C) and anti-cytochrome P450 1B1 (CYP1B1) (D) antibodies. Images were acquired at 20X magnification using a Leica Thunder microscope. Whole tissue slide images were mosaic merged via LAS-X software. (E) Representative images of immunofluorescence analysis of HIF-1α (green) and CYP1B1 (green) in two slides of ER-negative and positive breast cancer TMA. Immunofluorescence images were acquired using a Leica Thunder microscope. (F) Scatter plots showing fluorescence intensity correlation analysis of HIF-1α and CYP1B1 in 64 ER-positive TMAs. (G) Violin plots of fluorescence intensities of HIF-1α and CYP1B1. Samples were classified into low (immunohistochemical [IHC] score: 1+, 2+) and high (IHC score: 3+) groups according to the estrogen receptor (ER) expression status of the TMA specification sheet, and the correlation between HIF-1α and CYP1B1 expression was analyzed. (H) Analysis of fluorescence intensity of CYP1B1 against HIF1α expression in ER-positive TMA. The fluorescence intensity of CYP1B1 was analyzed by classifying the fluorescence intensity of HIF1-α based on the top and bottom 25% ER expression. *, P<0.0001 versus ER-positive tumor tissue with low HIF1-α fluorescence intensity by Mann-Whitney test.

Journal: American Journal of Cancer Research

Article Title: Upregulation of CYP1B1 by hypoxia is mediated by ERα activation in breast cancer cells

doi:

Figure Lengend Snippet: Analysis of HIF-1α and CYP1B1 expression in tissue microarray (TMA) samples from breast cancer patients. (A) Schematic diagram of the tumor stages of each breast tissue core. (B) Hematoxylin and eosin (H&E) staining images provided by the manufacturer. Immunofluorescence images of tumor microarrays (TMA) stained with anti-hypoxia-inducible factor-1 alpha (HIF-1α) (C) and anti-cytochrome P450 1B1 (CYP1B1) (D) antibodies. Images were acquired at 20X magnification using a Leica Thunder microscope. Whole tissue slide images were mosaic merged via LAS-X software. (E) Representative images of immunofluorescence analysis of HIF-1α (green) and CYP1B1 (green) in two slides of ER-negative and positive breast cancer TMA. Immunofluorescence images were acquired using a Leica Thunder microscope. (F) Scatter plots showing fluorescence intensity correlation analysis of HIF-1α and CYP1B1 in 64 ER-positive TMAs. (G) Violin plots of fluorescence intensities of HIF-1α and CYP1B1. Samples were classified into low (immunohistochemical [IHC] score: 1+, 2+) and high (IHC score: 3+) groups according to the estrogen receptor (ER) expression status of the TMA specification sheet, and the correlation between HIF-1α and CYP1B1 expression was analyzed. (H) Analysis of fluorescence intensity of CYP1B1 against HIF1α expression in ER-positive TMA. The fluorescence intensity of CYP1B1 was analyzed by classifying the fluorescence intensity of HIF1-α based on the top and bottom 25% ER expression. *, P<0.0001 versus ER-positive tumor tissue with low HIF1-α fluorescence intensity by Mann-Whitney test.

Article Snippet: A human breast cancer tissue microarray (TMA, #BC081120e) containing 110 cases was purchased from Biomax (Rockville, MD).

Techniques: Expressing, Microarray, Staining, Immunofluorescence, Microscopy, Software, Fluorescence, Immunohistochemical staining, MANN-WHITNEY