Journal: iScience

Article Title: From byproduct to biotherapeutic: Comparative study of buffy coats and leukoreduction system chambers for NK cell-based immunotherapies

doi: 10.1016/j.isci.2026.114907

Figure Lengend Snippet: Functional characterization of NK cells (A) Degranulation capacity of NK cells following a 2 h co-culture with K562 leukemia cells, measured by surface CD107a expression ( n = 5). (B and C) Europium-based cytotoxicity assay assessing the specific lysis of K562 target cells at E:T ratios of 10:1, 5:1, 3:1, 1:1, and 0.5:1 on day 7 (B) and day 14 (C) of culture to monitor functional decline over time ( n = 6 for both time points). Here, all comparisons were not significant and are summarized as such (n.s.: p > 0.05). (D) IFN-γ release following 16 h of co-culture of NK cells with K562 cells, quantified via Luminex assay ( n = 5). (E) Kinetic live-cell killing assay using the Incucyte monitoring of Nuclight Red signal loss in K562 cells over 72 h at an E:T ratio of 1:1 ( n = 6). (F) Proportion of donors reaching 25%, 50%, and 75% cumulative lysis thresholds during Incucyte co-culture with K562 cells. (G) CD107a degranulation assay of NK cells with or without pre-incubation with trastuzumab (anti-HER2 antibody) during a 2 h co-culture with MDA-MB-453 cells ( n = 5). (H) Incucyte-based cytotoxicity assay shows the real-time lysis of MDA-MB-453 cells by trastuzumab-loaded NK cells at an E:T ratio of 1:1 over 72 h ( n = 6). (I) Proportion of donors reaching 25%, 50%, and 75% cumulative lysis during the ADCC assay. (n varies as not all samples reached the benchmark values within the observation period). All ADCC experiments have been performed after 14 days of in vitro NK cell expansion. N/A indicates that statistical analysis was not possible for this group. 9J) CD107a degranulation of unmodified NK cells and ErbB2-CAR-NK cells after 2 h co-culture with MDA-MB-453 cells ( n = 5). (K) Incucyte-based cytotoxicity assay showing killing dynamics of ErbB2-CAR-NK cells against MDA-MB-453 cells at an E:T ratio of 1:1 over 72 h ( n = 6). (L) Proportion of donors reaching 25%, 50%, and 75% cumulative lysis during the CAR-mediated cytotoxicity assay. (n varies as not all samples reached the benchmark values within the observation period). All CAR-killing experiments have been performed after 14 days of in vitro NK cell expansion. Unless otherwise stated, n refers to biologically independent donor samples and is indicated per group. Data are shown as mean ± SEM. p-values were determined by Student’s t test with Welsh correction (A, D, F, G, I, J, and L) or by one-way ANOVA (B and C) with Tukey post-test (K and L).

Article Snippet: Mouse Monoclonal anti-CD340 (Her2) , Miltenyi Biotec , Cat# 130-124-474.

Techniques: Functional Assay, Co-Culture Assay, Expressing, Cytotoxicity Assay, Lysis, Luminex, Degranulation Assay, Incubation, ADCC Assay, In Vitro

Journal: eNeuro

Article Title: The Progestin Receptor Interactome in the Female Mouse Hypothalamus: Interactions with Synaptic Proteins Are Isoform Specific and Ligand Dependent

doi: 10.1523/ENEURO.0272-17.2017

Figure Lengend Snippet: List of antibodies used for RPPA analysis

Article Snippet: ErbB2/HER2(Y1248)_R_V , Imgenex , IMG-90189 , Rabbit.

Techniques: Plasmid Preparation

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: ACS Measurement Science Au

Article Title: Prototype Smartphone-Based Device for Flow Cytometry with Immunolabeling via Supra-nanoparticle Assemblies of Quantum Dots

doi: 10.1021/acsmeasuresciau.1c00033

Figure Lengend Snippet: Counting of SK-BR3 cells, stained with DAPI (cell nuclei) and immunolabeled with SiO 2 @(QD635-Dex)-(anti-HER2 TAC). (A) SK-BR3 cells only: (i–v) frames from smartphone FC videos of different concentrations of suspended cells. For the most dilute suspension, the arrow indicates a single cell. An intensity profile across selected cells can be found in Figure S13 . The image in (i) is a zoomed section of (v) with one (of several) SK-BR3 cells circled. Correlation plots (vi) of HER2-positive SK-BR3 cell counts derived from the smartphone FC versus a commercial cell counter. The diagonal line has a slope of unity. (B) Target SK-BR3 cells and background MDA-MB-231 cells: (i–v) frames from smartphone FC videos of mixed suspensions of SK-BR3 cells (variable amount) and MDA-MB-231 cells (approximately constant amount; DAPI stained). An intensity profile across selected cells of each type can be found in Figure S13 . Video S3 is a 10 s clip from one of the smartphone FC videos. The image in (i) is a zoomed section of (v) with one example each of a SK-BR3 cell and a MDA-MB-231 cell circled. Correlation plots of cell counts (vi) measured with a smartphone FC versus a commercial cell counter. Frame image brightness has been digitally increased for display purposes.

Article Snippet: Anti-HER2 antibody was from Novus Biologicals (Centennial, CO).

Techniques: Staining, Immunolabeling, Suspension, Derivative Assay

Journal: ACS Measurement Science Au

Article Title: Prototype Smartphone-Based Device for Flow Cytometry with Immunolabeling via Supra-nanoparticle Assemblies of Quantum Dots

doi: 10.1021/acsmeasuresciau.1c00033

Figure Lengend Snippet: Assessment of multiple colors of QDs. (A) PL emission spectra of QD490, QD540, QD585, QD605, QD635, and QD650 (solid lines). The approximate RGB color filter transmission spectra for the smartphone are overlaid (dashed lines; data courtesy of Olivier Burggraaff ). (B) Frames from smartphone FC videos showing SK-BR3 cells immunolabeled with SiO 2 @(QDλ-CM-Dex)-(anti-HER2) for QD540, QD585, QD605, or QD650. Frame image brightness has been digitally increased for display purposes. Intensity profiles across selected cells of each color of QD can be found in Figure S16 . (C) Plots of the G/B and R/B intensity ratios for individual SK-BR3 cells labeled with these four colors of SiO 2 @(QDλ-CM-Dex)-(anti-HER2). (D) Plots of the R, G, and B channel intensity values for individual SK-BR3 cells nonspecifically labeled with GSH-QD490, GSH-QD540, or GSH-QD635 (none as a SiO 2 @QD assembly).

Article Snippet: Anti-HER2 antibody was from Novus Biologicals (Centennial, CO).

Techniques: Transmission Assay, Immunolabeling, Labeling

Journal: ACS Measurement Science Au

Article Title: Prototype Smartphone-Based Device for Flow Cytometry with Immunolabeling via Supra-nanoparticle Assemblies of Quantum Dots

doi: 10.1021/acsmeasuresciau.1c00033

Figure Lengend Snippet: Counting of SK-BR3 and MDA-MB-231 cells in parallel using SiO 2 @(QD540-CM-Dex)-(anti-HER2) and SiO 2 @(QD650-CM-Dex)-(anti-MUC1), respectively. (A) Cartoon schematic of the experiment (not drawn to scale). (B) Example of a frame from a smartphone FC video. Frame image brightness has been digitally increased for display purposes. An intensity profile across selected cells of each type can be found in Figure S17 . Video S4 is a 20 s clip from one of the smartphone FC videos. (C) Examples of normalized G/B and R/B intensity ratios and cell classifications for five samples with an increasing number of MDA-MB-231 cells and an approximately constant number of SK-BR3 cells (see Figure S18 for additional data sets). Normalization of intensity ratios assisted with application of the SVM training set to the experimental data (see the SI for details). (D) Correlation plots for the measured and expected numbers of SK-BR3 and MDA-MB-231 cells, and for the measured and expected ratios of MDA-MB-231 and SK-BR3 cells. This plot includes the samples in panel (C) and in Figure S18 .

Article Snippet: Anti-HER2 antibody was from Novus Biologicals (Centennial, CO).

Techniques:

Journal: ACS Measurement Science Au

Article Title: Prototype Smartphone-Based Device for Flow Cytometry with Immunolabeling via Supra-nanoparticle Assemblies of Quantum Dots

doi: 10.1021/acsmeasuresciau.1c00033

Figure Lengend Snippet: Identification of selected breast cancer cell types (SK-BR3, MDA-MB-231, and MCF-7) using SiO 2 @(QD540-CM-Dex)-(anti-HER2), SiO 2 @(QD585-CM-Dex)-(anti-MUC1), and SiO 2 @(QD650-CM-Dex)-(anti-ER). (A) Cartoon schematic of the experiment (not drawn to scale). (B) Examples of G/B and R/B intensity ratios for a control sample without cells and suspensions of each cell type. The shaded region in the plots corresponds to the mean of the control sample data points plus four standard deviations along each intensity ratio axis. The circled regions highlight the dominant vertical, diagonal, and horizontal spread of cell counts for SK-BR3, MDA-MB-231, and MCF-7 cells, respectively.

Article Snippet: Anti-HER2 antibody was from Novus Biologicals (Centennial, CO).

Techniques: Control

Journal: Experimental and Therapeutic Medicine

Article Title: IL-35 improves T reg -mediated immune suppression in atherosclerotic mice

doi: 10.3892/etm.2016.3649

Figure Lengend Snippet: Plasma levels of Foxp3. Peripheral blood mononuclear cells were collected from each group, and the proportions of CD4 + CD25 + Foxp3 + T reg /CD4 + T-cells were analyzed by flow cytometry, and quantified using FlowJo 7.6.1 software. (A) Histogram. Data are presented as the mean ± standard deviation. *P<0.01, vs. the AS group. (B) Negative group, (C) AS group, (D) atorvastatin group and (E) IL-35 group. Foxp3, forkhead box protein 3; AS, atherosclerosis; IL-35, interleukin-35; APC, allophycocyanin; PE, phycoerythrin.

Article Snippet: Anti-Foxp3 antibody was purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China).

Techniques: Clinical Proteomics, Flow Cytometry, Software, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: IL-35 improves T reg -mediated immune suppression in atherosclerotic mice

doi: 10.3892/etm.2016.3649

Figure Lengend Snippet: Levels of Foxp3 in atherosclerotic lesions. The deposition of Foxp3 in arteries from various groups was detected by immunohistochemistry (magnification, ×400). Positive Foxp3 was shown as brown nuclei. In the (A) negative and (B) AS groups, the deposition of Foxp3 in the lesions was minimal. Conversely, Foxp3 deposition was increased in the lesions of the (C) atorvastatin and (D) IL-35 groups, as compared with the AS group. (E) This was shown to be significant following quantification. There was no significant difference in the levels of Foxp3 between the atorvastatin and IL-35 groups. *P<0.01, vs. the negative control. Foxp3, forkhead box protein 3; AS, atherosclerosis; IL-35, interleukin-35.

Article Snippet: Anti-Foxp3 antibody was purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China).

Techniques: Immunohistochemistry, Negative Control

Journal: International journal of oncology

Article Title: Although c‑MYC contributes to tamoxifen resistance, it improves cisplatin sensitivity in ER‑positive breast cancer.

doi: 10.3892/ijo.2020.4987

Figure Lengend Snippet: Figure 1. Establishment of TAM‑resistant breast cancer cells. (A) TAM‑resistant cells exhibited a more elongated, spindle‑shaped morphology compared with the respective parental cells. Magnification, x200. (B) Survival of TAM‑resistant cells and the respective parental cells was evaluated using a Cell Counting kit‑8 assay following treatment with various concentrations of TAM for 24 h. (C) Western blot analysis (D) and densitometry analysis of ERα and HER2 expression in MCF‑7, MCF‑7R, T47D and T47DR cells. Data are presented as the means ± standard deviation of the mean of 3 repeats. *P<0.05, **P<0.01, ***P<0.001 vs. respective parental cells. TAM, tamoxifen.

Article Snippet: Non‐specific binding sites were blocked by incubating the membranes with 5% non-fat milk, after which the membranes were incubated overnight at 4 ̊C with the primary antibodies: ERα (1:1,000); HER2 (1:1,000); AKT (1:1,000); p‐AKT (1:1,000); c‐MYC (1:1,000); cyclin D1 (1:1,000); vimentin (1:1,000); E‐cadherin (1:1,000); p21 (1:500); β‐catenin (1:500); GAPDH (1:1,000) Membranes were subsequently incubated with a horseradish peroxidase-conjugated anti-rabbit/mouse immunoglobulin G secondary antibody (1:1,000; cat. no. BA1075 and BA1051; Wuhan Boster Biological Technology, Ltd.).

Techniques: CCK-8 Assay, Western Blot, Expressing, Standard Deviation

Journal: Scientific Reports

Article Title: HLA-DR cancer cells expression correlates with T cell infiltration and is enriched in lung adenocarcinoma with indolent behavior

doi: 10.1038/s41598-021-93807-3

Figure Lengend Snippet: Mass cytometry antibody panel for lung adenocarcinoma.

Article Snippet: HER2 , 148Nd , Surface , 29D8 , Fluidigm , 3148011A.

Techniques: Mass Cytometry