Peer-reviewedCC BY-NC 4.0STAR Protocols
Protocol for live-cell imaging and analysis of NF-κB localization and target gene transcription in Drosophila S2∗ cells
- 1Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA
- 2Biological Design Center, Boston University, Boston, MA 02215, USA
- 3Department of Biology, Boston University, Boston, MA 02215, USA
Abstract
NF-κB transcription factor (TF) activation is heterogeneous across identically stimulated cells, shaping innate immune responses. Here, we present a protocol to quantify this heterogeneity in live Drosophila melanogaster S2∗ cells using dual reporters of NF-κB (Relish) subcellular localization and downstream transcription. We describe steps for reporter cell line generation, immune stimulation and time-lapse confocal imaging, and machine-learning-based single-cell analysis to classify signaling behaviors and predict transcriptional responses.
For complete details on the use and execution of this protocol, please refer to Nawar et al.1
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BibTeX
@article{nawar2026protocol,
title = {{Protocol for live-cell imaging and analysis of NF-κB localization and target gene transcription in Drosophila S2∗ cells}},
author = {Noshin Nawar and Emma Rits and Zeba Wunderlich},
journal = {STAR Protocols},
year = {2026},
month = oct,
doi = {10.1016/j.xpro.2026.104902},
url = {https://doi.org/10.1016/j.xpro.2026.104902},
note = {CC BY-NC 4.0},
}
RIS
TY - JOUR
TI - Protocol for live-cell imaging and analysis of NF-κB localization and target gene transcription in Drosophila S2∗ cells
AU - Noshin Nawar
AU - Emma Rits
AU - Zeba Wunderlich
PY - 2026
DA - 2026/10/20
JO - STAR Protocols
DO - 10.1016/j.xpro.2026.104902
UR - https://doi.org/10.1016/j.xpro.2026.104902
AB - NF-κB transcription factor (TF) activation is heterogeneous across identically stimulated cells, shaping innate immune responses. Here, we present a protocol to quantify this heterogeneity in live Drosophila melanogaster S2∗ cells using dual reporters of NF-κB (Relish) subcellular localization and downstream transcription. We describe steps for reporter cell line generation, immune stimulation and time-lapse confocal imaging, and machine-learning-based single-cell analysis to classify signaling behaviors and predict transcriptional responses. For complete details on the use and execution of this protocol, please refer to Nawar et al.1
SN - 2666-1667
ER -