Journal: International Journal of Molecular Sciences

Article Title: Effect of the Protein Corona Formation on Antibody Functionalized Liquid Lipid Nanocarriers

doi: 10.3390/ijms242316759

Figure Lengend Snippet: Schematic representation of the procedure steps for the preparation of LLCNs and subsequent functionalization to obtain immune-nanocapsules (LLNCs-αHER2).

Article Snippet: The monoclonal ErbB-2 antibody anti-human Vio ® Bright FITC (αHER2) (clone 24D2) from humans was obtained by Miltenyi Biotech (Madrid, Spain).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Effect of the Protein Corona Formation on Antibody Functionalized Liquid Lipid Nanocarriers

doi: 10.3390/ijms242316759

Figure Lengend Snippet: Mean diameter, standard deviation (SD), and mode of the LLNCs, LLNCs-αHER2, and LLNCs-αHER2-PC measured at 25 °C with NTA and DLS techniques in pH 7.4 buffer.

Article Snippet: The monoclonal ErbB-2 antibody anti-human Vio ® Bright FITC (αHER2) (clone 24D2) from humans was obtained by Miltenyi Biotech (Madrid, Spain).

Techniques: Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Effect of the Protein Corona Formation on Antibody Functionalized Liquid Lipid Nanocarriers

doi: 10.3390/ijms242316759

Figure Lengend Snippet: Hydrodynamic size distribution of the LLNCs, LLNCs-αHER2, and LLNCs-αHER2-PC measured at 25 °C with NTA technique in pH 7 buffer.

Article Snippet: The monoclonal ErbB-2 antibody anti-human Vio ® Bright FITC (αHER2) (clone 24D2) from humans was obtained by Miltenyi Biotech (Madrid, Spain).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Effect of the Protein Corona Formation on Antibody Functionalized Liquid Lipid Nanocarriers

doi: 10.3390/ijms242316759

Figure Lengend Snippet: Physico-chemical characterization: ( a ) Zeta potential of the LLNCs (▪), LLNCs-αHER2 ( ● ), LLNCs-αHER2-FBS ( ✱ ), and LLNCs-αHER2-PC ( ▷ ) measured at 25 °C as a function of the medium pH and low ionic strength. ( b ) SDS-PAGE analysis under reducing conditions of different LLNCs. (C) Molecular weight marker: (1) αHER2; (2) FBS; (3) Fibrinogen; (4) LLNCs-αHER2; (5) elution volume after cleaning LLNCs-αHER2; (6) LLNCs-αHER2-PC; (7) elution volume after cleaning LLNCs-αHER2-PC.

Article Snippet: The monoclonal ErbB-2 antibody anti-human Vio ® Bright FITC (αHER2) (clone 24D2) from humans was obtained by Miltenyi Biotech (Madrid, Spain).

Techniques: Zeta Potential Analyzer, SDS Page, Molecular Weight, Marker

Journal: International Journal of Molecular Sciences

Article Title: Effect of the Protein Corona Formation on Antibody Functionalized Liquid Lipid Nanocarriers

doi: 10.3390/ijms242316759

Figure Lengend Snippet: LLNCs-αHER2-PC after incubation of immune-nanocapsules in a simulated physiological medium with FBS supplemented with FB.

Article Snippet: The monoclonal ErbB-2 antibody anti-human Vio ® Bright FITC (αHER2) (clone 24D2) from humans was obtained by Miltenyi Biotech (Madrid, Spain).

Techniques: Incubation

Journal: International Journal of Molecular Sciences

Article Title: Effect of the Protein Corona Formation on Antibody Functionalized Liquid Lipid Nanocarriers

doi: 10.3390/ijms242316759

Figure Lengend Snippet: Representative confocal microscopy of ( A ) SKBR3 and ( B ) HDFa (scale bar = 20 μm) incubated for 60 min with NR-LLNCs, NR-LLNCs-HER2, and NR-LLNCs-HER2-PC. Red filter and green filter correspond to LLNCs labeled with Nile Red and LLNCs labeled with HER2-FITC, respectively.

Article Snippet: The monoclonal ErbB-2 antibody anti-human Vio ® Bright FITC (αHER2) (clone 24D2) from humans was obtained by Miltenyi Biotech (Madrid, Spain).

Techniques: Confocal Microscopy, Incubation, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Effect of the Protein Corona Formation on Antibody Functionalized Liquid Lipid Nanocarriers

doi: 10.3390/ijms242316759

Figure Lengend Snippet: In vitro uptake of NR-LLNCs, NR-LLNCs-HER2, and NR-LLNCs-HER2-PC in both SKBR3 and HDFa analyzed by flow cytometry. Values represent the fluorescence intensity of the merge channels for green and red as NR-LLNCs-HER2 are both red and green fluorescent. Data are mean values ± SD. * p < 0.001 shows the significant values calculated using t -test.

Article Snippet: The monoclonal ErbB-2 antibody anti-human Vio ® Bright FITC (αHER2) (clone 24D2) from humans was obtained by Miltenyi Biotech (Madrid, Spain).

Techniques: In Vitro, Flow Cytometry, Fluorescence

Journal: Nature Communications

Article Title: ERBB2 signaling drives immune cell evasion and resistance against immunotherapy in small cell lung cancer

doi: 10.1038/s41467-025-66800-x

Figure Lengend Snippet: a Relative phosphorylation of ERBB2 in primary and metastatic cells assessed by RTK-assay (n = 4 technical replicates). b Histogram showing ERBB2 expression determined by MFI in flow cytometry in representative human SCLC cell lines from primary tumor ( n = 1), pleural effusion ( n = 3), and liver metastases ( n = 2) with corresponding quantification ( n = 3 technical replicates). c Heat map of global protein profiling from primary and metastatic SCLC cells (each group n = 3 biological replicates). d Illustration of ERBB2 KO generation. Created in BioRender. Meder, L. (2025) https://BioRender.com/su67ybp . e Relative ERBB2 expression of WT and ERBB2 KO cells, determined by flow cytometry ( n = 4 biological replicates). f Relative MHC-I expression of WT and ERBB2 KO cells, determined by flow cytometry ( n = 4 biological replicates). g Relative B2M , H2-D1 expression in WT, ERBB2 KO and ERBB2 rescue in murine metastatic SCLC cell line measured by qPCR ( n = 3 biological replicates). h Heat map of RNA sequencing in WT and ERBB2 KO cells (each group n = 3 biological replicates). i Whole proteome analysis of WT and ERBB2 KO in murine metastatic SCLC cell line (each group n = 3 biological replicates). j Whole proteome analysis of murine metastatic SCLC cell line untreated vs. treated with neratinib/mubritinib (1 μM) for 24 h (each group n = 3 biological replicates). k Relative MHC-I expression after treatment with IFNγ (40 ng/mL) or IFNγ + ERBB2 inhibitor mubritinib (1 µM), determined by flow cytometry ( n = 3 biological replicates). l Relative MHC-I expression after treatment with IFNγ (40 ng/mL) or IFNγ + ERBB2 inhibitor mubritinib (1 µM), determined by flow cytometry ( n = 3 biological replicates). For flow cytometry analyses, MFI was normalized to IgG control and representative histograms are shown. Statistical analysis was performed using a two-sided, unpaired Student’s t -test. Data are represented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. Source data are provided as a Source Data file. Icons created in BioRender. Meder, L. (2025) https://BioRender.com/z2ehzfh .

Article Snippet: They were transfected with Ultracruz Transfection reagent (Santa Cruz, Cat. # sc395739) and the Plasmids containing Cas9 and the corresponding guide RNA (B2M Santa cruz, Cat. # sc-419281-nic or ERBB2 Santa cruz, Cat. # sc-420219-nic-2).

Techniques: Phospho-proteomics, Expressing, Flow Cytometry, RNA Sequencing, Control

Journal: Communications Biology

Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial–macrophage interaction

doi: 10.1038/s42003-025-08940-w

Figure Lengend Snippet: A Schemes for the irradiation site in the mouse lung and drug administration. C57BL/6 mice were irradiated in the left main bronchus with 90 Gy using a 4 mm diameter field, and EGFR TKIs (30 mg/kg) were administered every 2–3 d for a total of 2 weeks ( n = 5 animals per group). Mice were intraperitoneally injected with EGFR TKI and vehicle 1 h before irradiation. Representative images of the lungs; hematoxylin&eosin and Masson trichrome staining of lung tissues from C57BL/6 mice 2 weeks after irradiation with or without EGFR TKIs; scale bar = 100 µm. The fibrosis grade score is shown as an Ashcroft score graph. Collagen deposits were quantified using more than five fields (magnification: 200×). B HPAEps were treated with EGFR TKIs (1 μM) 1 h before treatment with the human-derived cytokines EGF, epiregulin, and NRG1 at 30 ng/mL for 5 min. Immunoblotting of EGFR, ERBB2, ERBB3, ERBB4, p-EGFR, p-ERBB2, p-ERBB3, p-ERBB4, and β-actin in HPAEps. The mean intensity of p-EGFR and p-ERBB3 from three independent experiments is given. In the Ashcroft score graph ( A ) and immunoblotting intensity graph ( B ), error bars indicate SD. In other graphs, error bars indicate SEM. Statistical significance was determined by one-way ANOVA with multiple comparisons (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant).

Article Snippet: Lipofectamine 2000 (Invitrogen; #11668019) was used as a transfection reagent, and EGFR siRNA (Santa Cruz Biotechnology; #sc-29301), ERBB2 siRNA (Santa Cruz Biotechnology; #sc-29405) ERBB3 siRNA (Santa Cruz Biotechnology; #sc-35327), ERBB4 siRNA (Santa Cruz Biotechnology; #sc-35329), STAT5 siRNA (Santa Cruz Biotechnology; #sc-29495), and control siRNA (Santa Cruz Biotechnology; #sc-37007) were transfected.

Techniques: Irradiation, Injection, Staining, Derivative Assay, Western Blot

Journal: Communications Biology

Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial–macrophage interaction

doi: 10.1038/s42003-025-08940-w

Figure Lengend Snippet: A C57BL/6 mice were irradiated in the left main bronchus with 90 Gy using a 4 mm diameter field. Lung samples were obtained before (0 month) and 8 and 14 d after irradiation and used for RNA-seq analysis. Representative images of hematoxylin&eosin staining of lung tissues from C57BL/6 mice 8, 11, and 14 d after irradiation; scale bar = 100 µm. Graphs quantifying collagen deposition per field (magnification, 200×). Heat map of RNA-seq analysis showing differentially expressed collagen genes in one of the three irradiated conditions. B EGFR/ERBB signaling genes regulated by irradiation in lungs. Differentially expressed genes 14 d after irradiation were visualized with EGFR inhibitors using Cytoscape. C Heatmap of RNA-seq analysis showing differentially expressed matrisome genes in one of the three irradiated conditions (8, 11, and 14 d). D Immunofluorescence staining of EGFR (green), ERBB3 (green), pro-SPC (red), and T1α (white) was performed on irradiated and control lung tissues; scale bar = 10 μm; scale bar of cropped images = 5 μm. The graphs show EGFR- and ERBB3-positive cells among T1α- and pro-SPC- positive cell populations. E Immunohistochemistry staining of p-EGFR, p-ERBB2, p-ERBR3, and p-ERBB4 in the lung tissues of control and C57BL/6 mice 2 weeks after irradiation ( n = 5 animals per group); scale bar = 20 µm; scale bar of cropped images = 5 µm. The average of five fields of p-EGFR, p-ERBB2, p-ERBB3, and p-ErbB4 density was quantified (magnification; 200×). In all graphs, statistical significance was determined by one-way ANOVA with multiple comparisons (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant).

Article Snippet: Lipofectamine 2000 (Invitrogen; #11668019) was used as a transfection reagent, and EGFR siRNA (Santa Cruz Biotechnology; #sc-29301), ERBB2 siRNA (Santa Cruz Biotechnology; #sc-29405) ERBB3 siRNA (Santa Cruz Biotechnology; #sc-35327), ERBB4 siRNA (Santa Cruz Biotechnology; #sc-35329), STAT5 siRNA (Santa Cruz Biotechnology; #sc-29495), and control siRNA (Santa Cruz Biotechnology; #sc-37007) were transfected.

Techniques: Irradiation, RNA Sequencing, Staining, Immunofluorescence, Control, Immunohistochemistry

Journal: Communications Biology

Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial–macrophage interaction

doi: 10.1038/s42003-025-08940-w

Figure Lengend Snippet: A Bleomycin (1.25 and 1.6 U/kg) in 50 μL PBS was injected intratracheally into mice. Micro-CT images show inflammatory- and fibrosis-phase mouse lungs after bleomycin administration. Masson’s trichrome staining and immunohistochemical detection of EGF, EREG, NRG1, and AREG in inflammatory- and fibrotic-phase mouse lungs after bleomycin administration; scale bar of trichrome image = 100 µm; scale bar of immunohistochemistry image = 2.5 µm. The density of EGF, EREG, NRG1, AREG, p-EGFR, p-ERBB3, and p-ERBB4 was quantified according to the Ashcroft score (magnification: 200×). B Mice were injected orally with EGFR TKIs (20 mg/kg) and nintedanib (60 mg/kg) 14 d after bleomycin administration. Drugs were administered orally daily for 2 weeks. Micro-CT images depict the lungs of mice after bleomycin injection after 2, 3, and 4 weeks in the fibrosis phase. The graphs show Hounsfield units (HU) for each group. C Hematoxylin&eosin and Masson’s trichrome staining of lung tissues from C57BL/6 mice 4 weeks after bleomycin injection treated with or without dacomitinib and nintedanib; scale bar = 100 µm. Fibrosis grade scoring and collagen deposition quantification per field (magnification: 200×) are shown. In the Ashcroft score graph ( C ), error bars indicate SD. Statistical significance was determined by one-way ANOVA with multiple comparisons (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant).

Article Snippet: Lipofectamine 2000 (Invitrogen; #11668019) was used as a transfection reagent, and EGFR siRNA (Santa Cruz Biotechnology; #sc-29301), ERBB2 siRNA (Santa Cruz Biotechnology; #sc-29405) ERBB3 siRNA (Santa Cruz Biotechnology; #sc-35327), ERBB4 siRNA (Santa Cruz Biotechnology; #sc-35329), STAT5 siRNA (Santa Cruz Biotechnology; #sc-29495), and control siRNA (Santa Cruz Biotechnology; #sc-37007) were transfected.

Techniques: Injection, Micro-CT, Staining, Immunohistochemical staining, Immunohistochemistry