Cooperation of kindlin-3 and talin-1 in integrin activation during neutrophil arrest and inflammation

Neutrophil arrest requires the activation of beta2 integrins. Two FERM domain proteins, kindlin-3 and talin-1, are necessary for neutrophil arrest. In this project, we explored the molecular mechanisms of talin-1 and kindlin-3 cooperation during neutrophil adhesion. Flow chamber experiments showed that talin-1 and kindlin-3 are simultaneously recruited to the plasma membrane of HL-60 cells during the transition from rolling to arrest. Using primary neutrophils expressing fluorescent fusion proteins of talin-1 or kindlin-3, we confirmed that both proteins are recruited to the plasma membrane at the time of arrest, supporting their coordinated role in neutrophil integrin activation. In a mouse cremaster muscle ischemia-reperfusion model, beta2 integrin activation was detected in neutrophils attached to the endothelium after ischemia-reperfusion. These findings support that kindlin-3 and talin-1 work together to induce high-affinity integrin activation during inflammation.
, HL-60 cells were differentiated for 5-7 days in 1.3 % DMSO and flowed on P-selectin and ICAM-1 substrate in a microfluidic device. Mouse neutrophils were isolated by flushing mouse tibia or femur. Cells were perfused at 6 dyn/cm2 wall shear stress through a microfluidic flow chamber and examined by total internal reflection fluorescence microscopy. Confocal microscopy was utilized to examine the antibody labelling in the cremaster muscle microcirculation of a male mouse before and after cremaster ischemia and reperfusion.
, # Cooperation of kindlin-3 and talin-1 in integrin activation during neutrophil arrest and inflammation

Dataset DOI: [10.5061/dryad.fttdz096g](https://doi.org/10.5061/dryad.fttdz096g)

## Description of the data and file structure

This dataset is associated with the American Heart Association (AHA) sponsored Career Development Award (942098) “Cooperation of kindlin-3 and talin-1 in integrin activation during neutrophil arrest and inflammation”. The Data_deposit.zip contains four experimental efforts, separated into subfolders:

1. The monitoring of the two cytosolic proteins talin-1 and kindlin-3 during HL-60 cell adhesion.

2. The monitoring of talin-1 during mouse neutrophil adhesion.

3. The monitoring of kindlin-3 during mouse neutrophil adhesion.

4. Confocal imaging of beta2 integrin activation in response to ischemia reperfusion injury (IRI). 

The dataset includes zipped files, which contain multiple files that need to be uncompressed. The folders are:

**Folder: HL-60.zip**
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