Additional file 3 of RBM45 associates with nuclear stress bodies and forms nuclear inclusions during chronic cellular stress and in neurodegenerative diseases

Additional file 3: Figure S3. Assessment of TDP-43 and FUS association with nuclear stress bodies (NSBs). (a) HEK293 cells were heat-shocked for 2 h at 42 °C to induce formation of NSBs. Cells were stained for endogeneous RBM45 and TDP-43 or FUS as indicated. Following heat shock, numerous RBM45-positive NSBs become visible in the cell nucleus (arrows) and these do not contain TDP-43 or FUS. Some TDP-43-positive stress granules are visible in the cytoplasm following heat shock and these do not contain RBM45. (b) HEK293 cells were transiently transfected to overexpress HA-tagged TDP-43 or FUS and were then stained for the HA tag and the NSB marker HSF1. Overexpression of TDP-43 was sufficient to induce NSB formation, but NSBs were negative for TDP-43. Neither overexpression of FUS or transfection with a control HA vector resulted in NSB formation. For all images, scale bar = 10 μm.