Fig. 3: ASK3 condensation is required for ASK3 inactivation under hyperosmotic stress. | Nature Communications

Fig. 3: ASK3 condensation is required for ASK3 inactivation under hyperosmotic stress.

From: Cells recognize osmotic stress through liquid–liquid phase separation lubricated with poly(ADP-ribose)

Fig. 3

a Schematic representation of ASK3 deletion mutants and fragments. The numbers indicate the amino acid (a.a.) positions in wild-type (WT). Black rectangle: kinase domain (652–908 a.a.), green rectangle: C-terminus coiled-coil domain (CCC: 1179–1225 a.a.), orange rectangle: C-terminus low-complexity region (CLCR: 1280–1293 a.a.). b, c Subcellular localization of ASK3 deletion mutants and fragments in HEK293A cells. Hyperosmotic stress: 600 mOsm, 10 min. DIC: differential interference contrast, white bar: 20 μm, red bar: 2.5 μm. A representative image set from four independent experiments is presented. Note that the signal intensity of DIC cannot be compared among the images. d Requirement of CCC and CLCR for ASK3 inactivation in HEK293A cells. Hyperosmolality (−): 300 mOsm; (+): 500 mOsm; 10 min. IB: immunoblotting, p-ASKendo: phosphorylation bands of endogenously expressed ASK. †Nonspecific bands. A representative image set from four independent experiments is presented (quantification: Supplementary Fig. 5a). e Relationship between ANKRD52 and ASK3 condensates in HEK293A cells. Magenta: ASK3-tdTomato, green: ANKRD52-Venus, hyperosmotic stress: 500 mOsm, white bar: 20 μm, yellow bar: 2.5 μm. A representative image set from five independent experiments is presented.

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