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. Author manuscript; available in PMC: 2024 Jun 1.
Published in final edited form as: Nat Chem Biol. 2023 Jun 22;19(12):1458–1468. doi: 10.1038/s41589-023-01353-y

Extended Data Fig.3. FAK-SPARK visualizes FAK activity with spatiotemporal resolution by targeting active FAK into specific locations.

Extended Data Fig.3

(A) Fluorescence images showing that FAK-SPARK visualizes FAK activity in the centrosome when constitutively active FAK (CA-FAK) is fused to aurora A kinase that is located in the centrosome (left panels). FAK activity is absent in the centrosome without CA-FAK (right panels). (B) Fluorescence images showing that nucleus-located FAK-SPARK-NLS (nuclear localization signal) visualizes FAK activity in the nuclear HOTag1 condensates that are tagged with CA-FAK (left panels). FAK activity is absent the CA-FAK-absent HOTag1 condensates. (C) Cartoon showing experimental procedure of measuring FAK-SPARK temporal resolution. (D) Time-lapse images after addition of rapamycin. (E) Normalized fluorescence along the dash line in (D). (F) Normalized SPARK signal over time. Scale bar, 5 μm (A, B, D).