Fig. 1. gp210-dsRNA-mediated depletion or mutation leads to formation of twinned nuclei. (A) Two-cell-stage embryos from control(RNAi), gp210(RNAi) or gp210(tm2320) nematodes were observed by DIC microscopy. Bar, 10 µm. (B) Total protein extracts from N2 (lane 1) or gp210(tm2320) (lane 2) nematodes were separated by SDS PAGE, transferred and probed with antibodies against gp210 or NUP107. (C) gp210 (green in merge) and NPCs labeled with mAb414 (red in merge) in fixed embryos expressing GFP-LEM2 (top) were observed by epifluorescence (dsRNA-treated embryos) or confocal microscopy [GFP::lem-2 and gp210(tm2320),GFP::lem-2 embryos)]. Bars, 10 µm. (D) gp210(RNAi) embryos were fixed by high-pressure freezing in utero and observed by TEM. Overview picture (left) of twinned nuclei and a higher magnification (right) of a normal NE containing NPCs (arrows). Note the presence of a stacked NE (arrowhead). Bars, 1 µm (overview) and 100 nm (zoom).