Have you been able to identify any nuclear pore complex biogenesis intermediates using this approach?

Hi,

That´s a very interesting possibility, but we have not been able to actually identify such intermediates probably due to a couple of reasons: 1) The nucleoporin we have mostly used as a handle for NPC purification is the nuclear basket component Mlp1, and the nuclear basket is probably one of the last NPC parts that gets assembled during its biogenesis; 2) Although we have been able to collect many thousands of particles for cryo-EM, it might still not be enough for the classification to identify intermediates that would represent only a small fraction and that could potentially not be easy to identify and align. Still, something to think about for the future and that could potentially be feasible using the right conditions.

Thanks for the answer.