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. 2013 Apr 27;41(12):6286–6299. doi: 10.1093/nar/gkt306

Figure 3.

Figure 3.

DDX3 drives the ATP-dependent assembly of gRNA translation granules. (A) Control (siCtrl) or DDX3-depleted (siDDX3) HeLa cells were transfected with 0.5 μg of pNL4-3 and 0.2 μg of pCIneo-HA-DDX3 (top and middle) or 0.2 μg of pCIneo-HA-DDX3R, which codes a siRNA-resistant DDX3 (bottom). At 24 hpt, cells were prepared for LSCM as indicated in ‘Materials and Methods’ section. Green staining corresponds to the gRNA, and red staining identifies DDX3. White arrowheads indicate cytoplasmic granules. Co-localized points are indicated in white. Merge images include nucleus staining with Hoechst (blue). Scale bars 25 μm (siCtrl/siDDX3) and 10 μm (siDDX3 + DDX3R). (B) HeLa cells were transfected with 0.5 μg of pNL4-3, 0.2 μg of pCIneo-HA-DDX3 (top) or pCIneo-HA-DDX3DQAD (middle) or pCIneo-HA-DDX3S382L (bottom) for 24 h and prepared for LSCM as indicated in ‘Materials and Methods’ section. Green staining corresponds to the gRNA, and red staining identifies DDX3. White arrowheads indicate cytoplasmic granules. Co-localized points are indicated in white. Merge images include nucleus staining with Hoechst (blue). Scale bars 10 μm (upper image) and 25 μm (middle and bottom images).