Supporting data pertaining to Nup60 phosphomutant localization and phosphorylation. (A, C, E, and G) Immunoblots of different Nup60 phosphomutants compared to Nup60-GFP (UB29253) in cdc20-mn background. Hxk2 was used as a loading control. The phosphomutants tested are: (A) Nup60^S89A-GFP (UB30327); (C) Nup60^Nterm3A-GFP (UB30329); (E) Nup60^Nterm5A-GFP (UB30331); and (G) Nup60^Cterm4A-GFP (UB30333). The Nup60-GFP control for A, C, and E are all reruns of the same replicate; the Nup60-GFP control for G is a rerun of the replicate from Fig. 5 G. The brackets to the left of the blots denote apparent phosphoshifts. (B, D, F, and H) Montages of cells with different fluorescently tagged Nup60 phosphomutants and Htb1-mCherry progressing through meiosis. The alleles visualized are: (B) Nup60^S89A-GFP (UB29265); (D) Nup60^Nterm3A-GFP (UB29441); (F) Nup60^Nterm5A-GFP (UB29443); and (H) Nup60^Cterm4A-GFP (UB29267). For B, D, F, and H, the white arrowheads in the “Fire LUT” images denote nuclei at the onset of anaphase I, the stage when Nup60-Nup2 detachment is observed. Scale bars, 2 μm. Source data are available for this figure: SourceData FS5.