Multimodal profiling reveals distinct endothelial activation pathways regulated by flow and heparan sulfate

This dataset contains the raw and processed quantitative outputs supporting the manuscript “Multimodal Profiling Reveals Distinct Endothelial Activation Pathways Regulated by Flow and Heparan Sulfate” (Harding, O’Hare, et al., 2025). The data quantify endothelial inflammatory and oxidative stress responses under four experimental conditions: (1) static, (2) uniform flow (12 dyn/cm²), (3) static + heparinase III, and (4) flow + heparinase III. The spreadsheets compile results from confocal immunofluorescence imaging, Western blot densitometry, reactive oxygen species (ROS) assays, and RNA sequencing. Quantified variables include mean fluorescence intensity (MFI) for protein markers (KLF2, ICAM-1, E-selectin, Nrf2, and Nf-κB p65), nuclear localization coefficients (Pearson’s coefficient for Nrf2 and DAPI), normalized band intensity ratios for Nox4, ICAM-1, and β-actin, and fluorescence-based ROS signals (H₂DCFDA and DHE assays). The RNA-seq dataset provides normalized counts, fold changes..., Overview
All numeric values included in the spreadsheets derive from image-based fluorescence analysis, Western blot densitometry, and RNA-seq differential-expression outputs. Each sample was classified by condition (Static, Flow, Static + Hep III, Flow + Hep III) and replicate number. No data were averaged or filtered beyond standard normalization steps, allowing full reuse for independent statistical analysis.

1. Image Quantification (Immunocytochemistry and ROS Assays)

Microscopy acquisition: Confocal (Zeiss LSM 710/800/880) images were collected using identical laser power, gain, and exposure settings within each experiment.

Mean Fluorescence Intensity (MFI): For each marker (KLF2, ICAM-1, E-selectin, Nrf2, Nf-κB), ImageJ/Fiji was used to select cell-level regions of interest. Background fluorescence was subtracted, and the integrated density was divided by area to yield MFI values.

Colocalization (Nrf2): Pearson’s correlation coefficient between Nrf2 and DAPI channels was calc..., # Multimodal profiling reveals distinct endothelial activation pathways regulated by flow and heparan sulfate

Dataset DOI: [10.5061/dryad.63xsj3vfg](https://doi.org/10.5061/dryad.63xsj3vfg)

## Description of the data and file structure

### Files and variables

#### File: Harding_OHare_Ebong_2025_Flow_HS_DataQuantification.xlsx

| <br /> | <br /> |
| :--------------------------------- | :------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ |
| **Sheet Name** | **Description** ...,