Fig. 3.

DP2392-E10 inhibits CRM1-mediated nuclear exports of NP-NES3 and NEP-NES2. (A) Schematic representation of Aequorea coerulescens green fluorescent protein (AcGFP)-NP-NES3, AcGFP-NEP-NES2, and AcGFP-Rev-NES constructs. (B-D) MDCK cells stably expressing AcGFP-NP-NES3, AcGFP-NEP-NES2, or AcGFP-Rev-NES were treated with DMSO, 10 nM LMB (positive control), or 2.5, 5.0, 7.5, 10.0, 12.5, or 15.0 µM DP2392-E10 for 8 h. The cells were then fixed with 4% paraformaldehyde, nuclei were stained with Hoechst 33342, and the mean fluorescent intensity of nuclear AcGFP was quantified on a CELAVIEW RS100. Data represents mean ±SD of nuclear AcGFP-NP-NES3 (B, bottom panel), AcGFP-NEP-NES2 (C, bottom panel), or AcGFP-Rev-NES (D, bottom panel) and the half-maximal effective concentration (EC₅₀) values from two independent experiments. Each of triplicate samples in one experiment was visualized for 36 fields of observation.