Effector-to-target Ratios
Effector-to-target Ratios - Related Videos
12:23
Analysis of Yersinia enterocolitica Effector Translocation into Host Cells Using Beta-lactamase Effector Fusions
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Cited by 5 •
2015
Effector translocation into host cells via a type III secretion system is a common virulence strategy among gram-negative bacteria. A beta-lactamase effector fusion based assay for quantitative analysis of translocation was applied. In Yersinia infected cells, conversion of a FRET reporter by the beta-lactamase is monitored using laser scanning microscopy.
08:39
Electroporation of Functional Bacterial Effectors into Mammalian Cells
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Cited by 7 •
2015
Electroporation was used to insert purified bacterial virulence effector proteins directly into living eukaryotic cells. Protein localization was monitored by confocal immunofluorescence microscopy. This method allows for studies on trafficking, function, and protein-protein interactions using active exogenous proteins, avoiding the need for heterologous expression in eukaryotic cells.
04:41
Introduction of Bacterial Effector Proteins into Mammalian Cells by Electroporation
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2025
In this video, we demonstrate the introduction of bacterial effector proteins into mammalian cells by electroporation. The intracellular localization of the bacterial effector proteins is detected by confocal microscopy following staining with specific fluorophore-conjugated antibodies.
02:12
Co-Infiltration Assay for Screening Pathogen Effectors that Suppress Plant RNA Silencing
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2026
Source: Shi, J., et al. Screening and Identification of RNA Silencing Suppressors from Secreted Effectors of Plant Pathogens. J. Vis. Exp. (2020)This video demonstrates a co-infiltration assay for identifying pathogen effector proteins that suppress RNA silencing in Nicotiana benthamiana 16c. A fluorescent reporter and a candidate effector gene are delivered into plant leaves via Agrobacterium. Normally, RNA silencing degrades reporter transcripts, reducing fluorescence. Suppressor effectors...
02:20
Monitoring Bacterial Infection and Effector Protein Delivery in Plant Cells
0 Views •
2025
Source: Lee, H. et. al., Split Green Fluorescent Protein System to Visualize Effectors Delivered from Bacteria During Infection. J. Vis. Exp. (2018)This video demonstrates a method for monitoring bacterial infection and effector protein delivery in plant cells using split superfolder green fluorescent protein (sfGFP) complementation combined with confocal microscopy. The approach enables precise visualization of effector translocation into the host cell cytosol.
FAQs
Changing the ratio alters the number of available effector cells relative to each target cell, which affects the likelihood and frequency of cell-cell contact during the assay. More frequent encounters create additional opportunities for recognition, immune synapse formation, and target-cell lysis within the incubation period. The ratio therefore connects cellular abundance with a measurable functional killing outcome.
A series of ratios tests how target-cell responses change as effector abundance changes. When targets show different levels of lysis across comparable ratio conditions, the pattern can reveal differences in susceptibility rather than simply indicating whether killing occurs. Researchers can use these comparisons to identify assay conditions that make variation among target cells more apparent.
The incubation period defines the time available for effectors and targets to encounter one another, undergo recognition, form immune synapses, and produce lysis. Consequently, a measured outcome reflects interactions occurring within that specified window, not an unlimited killing capacity. Keeping the period defined helps researchers interpret how the selected ratio relates to cytotoxic activity.
These ratios provide a common experimental basis for comparing immune-cell populations because each population can be examined at defined relative abundances. Differences in the resulting cytotoxic activity may indicate variation in the cells’ ability to contact, recognize, or lyse targets. The approach therefore supports functional comparison rather than relying only on cell identity or abundance.
Researchers select the effector and target cell populations, establish experimental conditions with different relative cell numbers, and incubate the combinations for a defined period. They then assess the resulting cytotoxic activity as a functional outcome and compare responses across ratios. This workflow shows how changing effector abundance influences interactions with the same type of target.
Testing a range of ratios can show whether measured cytotoxic activity changes as effector abundance increases or decreases. The resulting pattern helps researchers identify conditions that best reveal differences in target-cell susceptibility or immune-cell performance. It also supports comparisons among experiments by linking each functional outcome to a stated cellular relationship.
Effector-to-target measurements are useful in studies of host defense, infection, immunotherapy, and cancer. They connect immune-cell interactions with quantifiable functional outcomes, allowing researchers to examine cytotoxic activity in biologically relevant settings. In cancer and immunotherapy research, the approach can support comparisons of immune-cell populations and target susceptibility under defined experimental conditions.