Supporting data pertaining to a meiotic role for the Nup60 AH . (A) Immunoblots of Nup60-GFP (UB14646) and Nup60^ΔAH-GFP (UB25731) in cells progressing through meiosis. (B) Immunoblots of Nup60-GFP (UB14646) and Nup60^I36R-GFP (UB27189) in cells progressing through meiosis. For A and B, Hxk2 was used as a loading control and meiotic staging corresponding to the displayed blots was assessed using Htb1-mCherry. (C) Quantification of different Nup60 mutant protein levels during meiosis, corresponding to A and B. Background subtracted Nup60 intensity was divided by Hxk2 intensity to control for loading and then normalized to Nup60-GFP levels at 0 h in SPM (using the control run on the same blot). For Nup60-GFP, the mean ± standard deviation (shaded range) of four independent biological replicates is displayed. For Nup60^ΔAH-GFP and Nup60^I36R-GFP, the mean ± range (shaded area) of two independent biological replicates is displayed. (D and E) Montages of cells with either (D) Nup1-GFP (UB15303) or (E) Nup1^ΔAH-GFP (UB25727) and Htb1-mCherry going through meiosis. Schematics of NUP1 and nup1-ΔAH, which lacks its N-terminal AH, are displayed above their respective montages. For D and E, the white arrowheads in the “Fire LUT” images denote nuclei during meiosis I, a time point when Nup60^ΔAH detachment is apparent. (F) Montages of cells with Nup60^ΔAH-GFP and Htb1-mCherry exposed to different Cdc5 treatments in prophase I arrest (ndt80Δ): P_CUP1-CDC5^KD-3xFLAG-10xHis induced (UB29073), P_CUP1-CDC5-3xFLAG-10xHis uninduced (UB29071), and P_CUP1-CDC5-3xFLAG-10xHis induced (UB29071). Induction of Cdc5 protein was performed at 5 h in SPM with 50 μM CuSO₄. (G) Quantification of Nup60^ΔAH detachment for the experiment depicted in F. Individual DI values were normalized to the average DI for uninduced P_CUP1-CDC5^KD-3xFLAG-10xHis cells at the pre-induction time point (5 h). Asterisks indicate statistical significance calculated using a Wilcoxon signed-rank test when post-induction (6 h in SPM) values were compared to preinduction (5 h in SPM) values for each treatment regimen (see Table S5 for P values). Sample sizes (n) are the number of cells quantified for each treatment regimen. (H) Immunoblots for Nup60^ΔAH-GFP and Cdc5^KD-3xFLAG-10xHis (UB29073) or Cdc5-3xFLAG-10xHis (UB29071) before (5 h in SPM) or after (6 h in SPM) treatment (either addition of copper or not) during prophase I arrest (ndt80Δ), corresponding to the images in F. Hxk2 was used as a loading control. The bracket to the left of the blot denotes the apparent phosphoshift. For each montage, normalized DI values are indicated when calculated. Scale bars, 2 μm. Source data are available for this figure: SourceData FS6.