JCS009225 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Overexpressed MLN51 can be found in cytoplasmic granules positive for stress-granule (SG)-resident proteins. (A) Immunofluorescence of HeLa cells transiently transfected using pEYFP-MLN51/1-703 plasmid. Cells were directly observed after fixation. Note that MLN51 shows either a diffuse (a) or punctate (b,c) cytoplasmic staining. (B) MLN51, FMRP and PABP colocalize in cytoplasmic foci of HeLa cells transiently transfected with the pEYFP-MLN51/1-703 plasmid. Cells were fixed, permeabilized and incubated with an anti-FMRP (a,d, red), anti-PABP (e,h, red) or anti-Dcp1 (I,l, red) antibody. Note that cytoplasmic foci positive for MLN51 are distinct from Dcp bodies but both structures are occasionally very close to each other (arrowheads). Nuclei were counterstained with Hoechst 33258 dye (blue). (C) Transfection can induce SG formation. Immunofluorescence of HeLa cells transiently transfected using pEYFP plasmid expressing EYFP alone (green). Cells were fixed and stained with anti-FMRP (red). Note that a transfected cell with EYFP alone shows a punctate staining of FMRP while untransfected cells shows a diffuse cytoplasmic staining.

• Supplemental Figure S2 -

  Fig. S2. Nocodazole prevents stress granule (SG) assembly. Immunofluorescence analysis of HeLa cells untreated or treated with arsenite in the presence or in the absence of nocodazole. Cells were fixed and stained with anti-MLN51 (red) and anti-tubulin (green) antibodies. Nuclei were counterstained with Hoechst 33258 dye (blue). Note that in the presence of both arsenite and nocodazole, no MLN51-positive cytoplasmic foci is present and tubulin is depolymerized.