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For experiments, cells were used at passages 5–10.
In iCasper experiments, cells were examined by laser confocal microscope (FV1200, Olympus) under × 200 magnification.
For inhibitor experiments, cells were pre-treated with the inhibitor for one hour prior to stimulation.
For Ussing chamber experiments, cells were plated onto Costar snapwell inserts (12 mm), as previously described14.
For imaging experiments, cells were plated on coverglass-bottomed dishes (In Vitro Scientific).
In control experiments, cells without added polyQ40 show basal levels of aggregation (Figure 1b).
In a subset of experiments, cells were also cultured with 10 μM Alk5i.
For routine expansion and further experiments, cells were grown in growth media supplemented with laminin instead of coating the dishes.
In all T-cell genome editing experiments, cells were incubated at 30 °C for 40 h following nanoparticle transfection.
For fluorescence loss in photobleaching (FLIP) experiments, cells were photobleached every frame until the end of the imaging.
For SNX2 rescue experiments, cells were transfected with plasmid DNA 10 h after the second siRNA transfection.
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