Journal: PLoS ONE

Article Title: A Simple and Sensitive Method for Measuring Tumor-Specific T Cell Cytotoxicity

doi: 10.1371/journal.pone.0011867

Figure Lengend Snippet: 4T1-Her2 cells were seeded in 96 well plates at increasing numbers and the luciferase level was determined after 2 hours. Both cell number and luciferase quantity were converted into percentage by dividing the individual figure with the maximal cell number seeded (100,000) or the luciferase reading from the well with the maximal cell number. The correlation coefficient (R) is indicated in the figure.

Article Snippet: The sorted GFP positive cells were then seeded as single cells in 96-well plate by limiting dilution and screened for colonies expressing HER2 by staining them with PE conjugated -anti-human HER2 antibody (BioLegend, San Diego, CA).

Techniques: Luciferase

Journal: PLoS ONE

Article Title: A Simple and Sensitive Method for Measuring Tumor-Specific T Cell Cytotoxicity

doi: 10.1371/journal.pone.0011867

Figure Lengend Snippet: Initially 10,000 4T1-Her2 cells were seeded. The supernatants were collected at the indicated times for measurement of luciferase.

Article Snippet: The sorted GFP positive cells were then seeded as single cells in 96-well plate by limiting dilution and screened for colonies expressing HER2 by staining them with PE conjugated -anti-human HER2 antibody (BioLegend, San Diego, CA).

Techniques: Luciferase

Journal: PLoS ONE

Article Title: A Simple and Sensitive Method for Measuring Tumor-Specific T Cell Cytotoxicity

doi: 10.1371/journal.pone.0011867

Figure Lengend Snippet: A . Cell-associated and supernatant luciferase activity during cTCR-splenocyte mediated cytolysis of 4T1-Her2 cells. cTCR-splenocytes were added to 4T1-Her2 cells at a ratio of 10∶1. Supernatants and cells were harvested at the indicated time for quantification of luciferase activity. B . 4T1-Her2 monolayer without cTCR-transduced splenocytes. C . Seventy-two h after cTCR-tranduced splenocytes were added to 4T1-Her2 monolayer.

Article Snippet: The sorted GFP positive cells were then seeded as single cells in 96-well plate by limiting dilution and screened for colonies expressing HER2 by staining them with PE conjugated -anti-human HER2 antibody (BioLegend, San Diego, CA).

Techniques: Luciferase, Activity Assay

Journal: PLoS ONE

Article Title: A Simple and Sensitive Method for Measuring Tumor-Specific T Cell Cytotoxicity

doi: 10.1371/journal.pone.0011867

Figure Lengend Snippet: Mock-transduced ( A ) or cTCR-transduced ( B ) splenocytes were added to 4T1-Her2 cells at the indicated ratio. Cells were harvested 72 later for quantification of luciferase activity. C . Using the formula: % specific luc reduction = (% luc reduction from engrafted T-cell)/(% luc reduction from control T-cell)×100, the data from A and B were converted into percentage of specific luciferase release and plotted.

Article Snippet: The sorted GFP positive cells were then seeded as single cells in 96-well plate by limiting dilution and screened for colonies expressing HER2 by staining them with PE conjugated -anti-human HER2 antibody (BioLegend, San Diego, CA).

Techniques: Luciferase, Activity Assay, Control

Journal: Cell Reports Methods

Article Title: Sequence enrichment profiles enable target-agnostic antibody generation for a broad range of antigens

doi: 10.1016/j.crmeth.2023.100475

Figure Lengend Snippet:

Article Snippet: HER2 , Sino Biological , Cat# 10004-H08H.

Techniques: Produced, Recombinant, Sequencing, Software

Journal: Biochemical pharmacology

Article Title: BRET-based assay to monitor EGFR transactivation by the AT 1 R reveals G q/11 protein-independent activation and AT 1 R-EGFR complexes

doi: 10.1016/j.bcp.2018.10.017

Figure Lengend Snippet: (A) Schematic of AT1R-EGFR BRET-based transactivation. EGFR fused to a BRET donor (Rluc8), is co-transfected with Grb2 adaptor protein tagged with a BRET acceptor (Venus) and the AT1R. Stimulation of the AT1R promotes activation of the EGFR and recruitment of Grb2. (B) HEK293 cells expressing AT1R, EGFR-Rluc8, and Grb2-Venus were treated with 10μM AngII, 1μM EGF or vehicle. Quantification of ligand-induced BRET ratio (maximum-minimum) between EGFR-Rluc8 and Grb2-Venus following AngII- and EGF-stimulation. Insert is HEK293 cells (stably expressing AT1R) stimulated with 100nM AngII, 10nM EGF or vehicle for 5 minutes before processing for phospho-ERK1/2:total-ERK1/2 (p-ERK:T-ERK) western blots. Blots are representative of 3 independent experiments (B inset) Cells expressing AT1R, Grb2-Venus and either EGFR-Rluc8, HER2-Rluc8 or HER3-Rluc8 and stimulated with 10μM AngII. Agonist stimulation is indicated by arrow. Data represent mean ± SEM of 3 independent experiments.

Article Snippet: EGFR-Rluc8, HER2-Rluc8 and HER3-Rluc8 were generated by inserting EGFR, HER2, and HER3, obtained from Origene (Rockville, MD, USA) into pcDNA3-Rluc8.

Techniques: Transfection, Activation Assay, Expressing, Stable Transfection, Western Blot