Nsp1-GFPCYT foci interact with ER in the daughter cell. In all images shown, cdc6-1 cells were arrested in G2/M for 3 h at 34°C and then imaged at RT. (A–C) Deconvolved fluorescence micrographs of one z section of cdc6-1 cells expressing Nsp1-GFP and either Kar9-mCherry (PCCPL535), Exo70-mCherry (PCCPL532), or HDEL-DsRed (PCCPL533). Green and red channels are shown in addition to the merge. Arrowheads point to Nsp1-GFPCYT foci. (D) A time-lapse series (Δt = 5 s) of PCCPL533 cells. Each time point is a merge of Nsp1-GFP and HDEL-DsRed images from one z plane. Arrowheads point to a Nsp1-GFPCYT focus (Video 4). (E) PCCPL533 cells were treated with latrunculin A (lat A), nocodazole, or DMSO. Deconvolved micrographs from one z section of a representative cell are shown (green, red, and merged images). Nsp1-GFPCYT foci are indicated by arrowheads. (F) Quantification of E showing the distribution of Nsp1-GFPCYT between the cortical and tubular ER under the indicated conditions. 58 ≤ n ≤ 71. Bars, 2 µm.
