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. 2022 May 12;13:2638. doi: 10.1038/s41467-022-30303-w

Fig. 3. The IDR1 is required for MRNIP phase separation.

Fig. 3

a The disordered region of MRNIP was analysed using PONDR. b Schematic of the optoIDR assay. Cells expressing recombinant protein with an IDR, mCherry and Cry2 were exposed to blue light (488 nm). c IDR1- or IDR2- Cry2-mCherry was expressed in cells, which were stimulated with blue light to induce condensation. d FRAP of blue light-induced IDR1-Cry2-mCherry droplets. n = 3 foci analysed in 3 independent experiments. Data are presented as means ± SEM. e, f Blue light-induced IDR1-Cry2-mCherry droplets were sensitive to 10% 1,6-hexanediol. g IDR1-Cry2-mCherry droplets fused to form a larger droplet upon stimulation with blue light. h-i IDR1-deleted MRNIP could not form liquid-like droplets in vivo (h) or in vitro (i). The droplets formed in a buffer described in Fig. 2n and a protein concentration of 10 μM was used in (i). For (c–h), HeLa cells were transfected with plasmid for 24 h before observation using confocal microscopy.