Fig. 8: Overexpression of Apq12 restores the Vps4 localization in lem2Δlnp1Δ cells during interphase.
From: Lem2 and Lnp1 maintain the membrane boundary between the nuclear envelope and endoplasmic reticulum
a Localization of Vps4 and Cmp7. Vps4-mCherry (magenta) and Cmp7-GFP (green) were expressed in Apq12-overexpressing lem2Δlnp1Δ cells. Projected images after deconvolution are shown. White lines indicate the outline of the cell. BF represents bright field image. Scale bar represents 5 μm. b, c Quantification of the number of Vps4 and Cmp7 dots. The number of Vps4 (b) and Cmp7 (c) dots per cell were counted and plotted as a box-and-whisker plot: horizontal lines in the box indicate the upper quartile, median, and lower quartile, from top to bottom, respectively; the whisker indicates standard deviation. The data for WT and lem2Δlnp1Δ cells are reproduced from Fig. 6g, h for comparison. n indicates the total cell number counted. p Values are from Steel–Dwass test. ***p < 0.001, n.s.: no significance (p > 0.05). d Protein levels of Vps4. The WT and Apq12-overexpressing lem2Δ (left panels) and lem2Δlnp1Δ cells (right panels) were subjected to SDS-PAGE. The protein levels of Vps4 and Cmp7 were detected as described in Fig. 6b. The full, uncropped blot images are shown in Supplementary Fig. 14. e Mitotic localization of Vps4 in Apq12-overexpressing lem2Δlnp1Δ cells. Vps4-GFP (green) was co-expressed with an SPB marker Sfi1-mCherry (magenta) in Apq12-overexpressing lem2Δlnp1Δ cells and observed in live. Insets on the right represent enlarged images of the arrowed SPB. Scale bar represents 5 μm. f Genetic interactions in maintaining the NE–ER boundary.
