Fig. 3.

ELYS is required for proper nuclear pore assembly. Immunofluorescence on HeLa cells transfected with control, ELYS, or Nup133 siRNA duplexes for 48–60 h. Cells were Triton X-100-extracted and then fixed and stained with the antibodies shown. Arrows indicate transfected cells. (A) ELYS RNAi results in a knockdown of ELYS and mislocalization of Nup133, whereas Nup133 RNAi similarly results in a knockdown of Nup133 and a reduction of ELYS in the nuclear envelope. ELYS RNAi leads to the mislocalization of the FG-Nups (mAb414), Nup160, and Nup358 from the nuclear rim to cytoplasmic aggregates (center column). (B) A magnification of ELYS-depleted cells clearly shows that Nup133 is greatly reduced in nuclear envelope pores and is mislocalized to cytoplasmic aggregates. (C and D) ELYS RNAi did not significantly affect lamin A/C localization to the nuclear lamina (C), or the nuclear accumulation of the transport factor Ran (fixed before permeabilization) (D). (Scale bars: 10 μm.)