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. 2018 Oct 1;32(19-20):1321–1331. doi: 10.1101/gad.315523.118

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© 2018 McCloskey et al.; Published by Cold Spring Harbor Laboratory Press

This article, published in Genes & Development, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.

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Figure 5. — Tpr regulates NPC number at the NE. Mean values of NPC numbers are indicated by red bars. t-test was performed to obtain P-value. (***) P < 0.0001. More than 20 nuclei were used to obtain the total NPC number or the density in each condition. (A) The localization of siRNA-resistant Tpr constructs was investigated by immunofluorescence using Flag tag. The dN mutant, which carries two point mutations in the Nup153-binding site (L458D and M489D), cannot localize at the NE due to the lack of Nup153-binding ability. (B,C) siRNA against endogenous Tpr was transfected to U2OS cells, which stably expressed the siRNA-resistant Tpr mutant. Seventy-two hours after transfection, total NPC number and the density were obtained by counting Nup153 foci using SR-SIM.