Journal: Nature Communications

Article Title: Two-input protein logic gate for computation in living cells

doi: 10.1038/s41467-021-26937-x

Figure Lengend Snippet: a Images of rapamycin-treated Ch -FAK-expressing FAK −/− fibroblasts treated with control (DMSO) and FAK inhibitor 14. Top panel shows the Ch -FAK-activated FAK −/− cells treated with control. Arrows indicate focal adhesions. Bottom panel shows same set of cells treated with FAK inhibitor 14. Scale bar, 20 μm. b Images of Ch- FAK-expressing FAK −/− fibroblasts without (top) and with (bottom) rapamycin stained with phalloidin to reveal stress fibers (magenta). Large focal adhesions formed by the activation of Ch -FAK are visible as red clusters (mCherry fluorescence) at the ends of stress fibers in bottom panel. Nuclei were stained with Hoechst (blue). Individual channels are shown to the right of the merged images. Scale bar, 20 μm. c Images of rapamycin-treated Ch -FAK-expressing FAK −/− fibroblasts treated with control (DMSO) (top) or blebbistatin (bottom). Red indicates phalloidin stained stress fibers and green indicates Hoechst-stained nucleus. Scale bar, 40 μm.

Article Snippet: We stained F-actin with Alexa Fluor phalloidin conjugates (ThermoFisher Scientific; 10 U/mL in 1% BSA PBS) and chromatin with 1:1000 Hoechst solution (Tocris).

Techniques: Expressing, Control, Staining, Activation Assay, Fluorescence