Fig. 2. Acute gain- and loss-of-function approaches support a role for fractionally induced effectors of nucleocytoplasmic transport imperceptible at the population level.

a–d Candidates related to nucleocytoplasmic transport affect DE penetrance induced by ErbB heterodimerization when ectopically expressed. B2B1 cells expressing inducible FLAG-tagged CSE1L (a), NPIPB11 (b), NUP37 (c), or KPNB1 (d) were used where indicated. e, f Loss of ErbB heterodimer-induced DEs upon inducible knockdown of nucleocytoplasmic regulators. B2B1 cells expressing inducible shCSE1L (e) or shNUP37 (f) were used where indicated. g–j Quantitative PCR for CSE1L (g), NPIPB11 (h), NUP37 (i), and KPNB1 (j) in B2B1 cells cultured in 3D for 6 days followed by addition of 0.5 µM AP21967 (AP) for 24 h or 72 h where indicated. Data are shown as the geometric mean (normalized to the 24-h, minus-AP condition) ± log-transformed s.e. from n = 7 (plus-AP, 24 h) or 8 biological replicates (all other groups). Differences in geometric means were assessed by two-sided t test after log transformation. For a–f, B2B1 cells stably expressing doxycycline (DOX)-inducible ectopic constructs were 3D cultured for 9–13 days with or without 0.5 µM AP added at Day 6 and/or 1 µg/ml DOX added at Day 5 where indicated. Data are shown as the arcsine transformed mean ± s.e. from n = 4 biological replicates where >100 outgrowths were scored per replicate. Differences by factor (DOX or AP) and two-factor interaction (int) were assessed by two-way ANOVA after arcsine transformation. Ectopic expression was confirmed by immunoblotting for FLAG with tubulin used as a loading control, except for NPIPB11 where induction was confirmed by quantitative PCR. Inducible knockdown was quantified relative to the no DOX shRNA control targeting luciferase (shLuc). Source data are provided as a Source Data file.