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TfR1 and <t>EGFR</t> expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and <t>anti-EGFR</t> micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).
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TfR1 and <t>EGFR</t> expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and <t>anti-EGFR</t> micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).
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TfR1 and <t>EGFR</t> expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and <t>anti-EGFR</t> micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).
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TfR1 and <t>EGFR</t> expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and <t>anti-EGFR</t> micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).
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TfR1 and <t>EGFR</t> expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and <t>anti-EGFR</t> micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).
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TfR1 and <t>EGFR</t> expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and <t>anti-EGFR</t> micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).
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TfR1 and <t>EGFR</t> expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and <t>anti-EGFR</t> micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).
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egfr (d38b1; #4267) antibody  (Cell Signaling Technology Inc)
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TfR1 and <t>EGFR</t> expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and <t>anti-EGFR</t> micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).
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Image Search Results


TfR1 and EGFR expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and anti-EGFR micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).

Journal: ACS Applied Materials & Interfaces

Article Title: Dual-Targeting Strategy to Repurpose Cetuximab with HFn Nanoconjugates for Immunotherapy of Triple-Negative Breast Cancer

doi: 10.1021/acsami.5c06626

Figure Lengend Snippet: TfR1 and EGFR expression in U-87 MG and MDA-MB-231 cells. (A) Representative Western blot of total lysates of U-87 MG and MDA-MB-231 cells (15 μg) blotted as indicated. (B) Densitometric quantification of the TfR1 and EGFR signal normalized on the corresponding β-actin signal. Bar graphs show mean ± s.e. of 3 independent replicates. ** = p < 0.01 (two-tailed Student’s t -test). (C) Quantification of the fluorescence intensity of HFn alone or conjugated with CTX normalized by the number of nuclei. Data were obtained by measuring HFn–AF647 fluorescence in U-87 MG and MDA-MB-231 cells after 2 min of incubation with HFn or HFn–CTX. (D) Cell-surface EGFR and TfR1 levels as measured with CellProfiler and representative images of non-permeabilized cells stained for EGFR or TfR1. Nuclei were stained with Hoechst (blue), TfR1 and EGFR (red), and CFDA SE-labeled MDA-MB-231 cells (green). White asterisks mark the position of the MDA-MB-231 cells in the anti-TfR1 and anti-EGFR micrographs. Scale bar, 20 μm. Violin superplots show the mean ± s.e. of 3 independent replicates (1665 U-87 MG cells and 1065 MDA-MB-231 cells for EGFR; 2178 U-87 MG cells and 1751 MDA-MB-231 cells for TfR1). ** = p < 0.01; **** p < 0.0001 (unpaired two-tailed t -test).

Article Snippet: Primary antibodies: CD44 (clone IM7; Bio-Rad, MCA4703), CD31/PECAM-1 (Biotechne, AF3628), CD71 (clone D7G9X, Cell Signaling, 13113), , EGFR (clone D38B1, Cell Signaling, 4267), β-Actin (clone D6A8, Cell Signaling, 8457), EGFR (clone 528, Merck, GR01), CD71 (clone MEM-75, Invitrogen, MA1-19137), and Ferritin Heavy Chain (clone EPR3005Y, Abcam, ab75972).

Techniques: Expressing, Western Blot, Two Tailed Test, Fluorescence, Incubation, Staining, Labeling