Fig. 3

Domains required for RBM45 incorporation into nuclear stress bodies (NSBs). a Schematic showing functional domains and their position in the full-length RBM45 protein. RRM = RNA recognition motif, HOA = homo-oligomer assembly domain, NLS = nuclear localization sequence. HEK293 cells were transfected with constructs encoding N-terminally HA-tagged wild-type (WT) or domain-modified forms of RBM45 as indicated to determine which domains of the protein are necessary for incorporation into NSBs. b In unstressed cells, the distribution of WT RBM45 is diffuse and nuclear and no NSBs are observed. c Heat shock (42 °C for 2 h) leads to the robust formation of NSBs that are positive for wild-type (WT) RBM45 and the NSB marker HSF1. d Removal of the RBM45 NLS (∆NLS) leads to its sequestration in the cytoplasm and prevents its association with NSBs during heat shock. e Removal of RRM1 (∆RRM1) does not alter the association of RBM45 with NSBs. f Removal of RRM2 (∆RRM2) prevents the association of RBM45 with NSBs. g Removal of RRM3 (∆RRM3) prevents the association of RBM45 with NSBs. h Removal of the RBM45 HOA domain (∆HOA) does not alter the association of RBM45 with NSBs. For all images, scale bar = 5 μm