Fig. 6: In vitro liquid–liquid phase separation of SAP97/hDLG.
From: Nanoscale regulation of Ca2+ dependent phase transitions and real-time dynamics of SAP97/hDLG
A A scheme indicating purified SH3-HOOK-GUK isoforms, namely I2 and I3, of C-terminal fragments of SAP97/hDLG and Calmodulin flanked by enhanced green fluorescent protein and monomeric mCherry, respectively, at the C-terminus. B The electrostatic distribution of net surface charges of SH3-I2-GUK, SH3-I3-GUK and Calmodulin. Pseudo color scalebar indicates transition from positive to negative (red to blue). C Top and bottom panels indicate the phase separation of C-terminal isoforms, namely SH3-I2-GUK::GFP and SH3-I3-GUK::GFP in response to their increasing concentration at 20% PEG. SH3-I2-GUK::GFP nucleation occurs at a concentration higher than SH3-I3-GUK::GFP, indicating that spontaneous phase transition of I3 isoform of SAP97/hDLG occurs at a lower concentration than that of the I2 isoform. Scalebar indicates 5 µm. D Incubation of 2 µM of CaM::mCherry with 1 µM SH3-I2-GUK::GFP or SH3-I3-GUK::GFP in presence of 2 mM CaCl2 resulted in the formation of co-condensates of SAP97/hDLG and Calmodulin. In the overlap images, SAP97/hDLG is in magenta and Calmodulin in green and their overlay is indicated by black. Scalebar indicates 3 µm.
