Skip to main content
. 2021 Nov 24;11(11):210250. doi: 10.1098/rsob.210250

Figure 4.

Figure 4.

Overexpression of APQ12 causes the mislocalization of ER proteins. (a) Overexpression of APQ12 is toxic for the cells. WT cells with the vector control, PGal1-APQ12 or PGal1-APQ12-6His were spotted in 10-fold serial dilutions onto YPAD (Glu) and YARaf/Gal (Gal/Raf) plates at 30°C. (b) Of the APQ12, BRL1, BRR6 module only APQ12 overexpression is toxic. WT cells with the vector control, PGal1-APQ12, PGal1-BRR6 and PGal1-BRL1 were spotted in 10-fold serial dilutions onto YPAD (Glu) and YARaf/Gal (Gal/Raf) plates at 30°C. (c) Overexpression of APQ12 causes mislocalization of yeGFP-Brl1. Cells with either the vector control (electronic supplementary material, figure S3C) or PGal1-APQ12-6His were incubated with galactose for the indicated times. The boxed cell at 1 h is a two-fold enlargement of the selected cell. (d) Line scan along the NE of a PGal1-APQ12-6His yeGFP-BRL1 NUP85-tdTomato cell (enlarged boxed cell in (c)) incubated for 1 h with galactose. It shows that yeGFP-Brl1 and Nup85-tdTomato are localized on separate domains along the NE. (e) Immunoblot of PGal1-APQ12-6His yeGFP-BRL1 NUP85-tdTomato cells. The pGal1 promoter was induced by the addition of galactose (t = 0). Samples were taken after the indicated times. Tub2 is a loading control. Apq12-6His was detected by anti-His antibodies. (f) Overexpression of APQ12 causes mislocalization of yeGFP-Brr6 and Nup85-tdTomato. Cells with either the vector control (electronic supplementary material, figure S3D) or PGal1-APQ12-6His were incubated with galactose for the indicated times. The boxed cell at 1 h is a twofold enlargement of the selected cell. (g) Line scan along the NE of a PGal1-APQ12-6His yeGFP-BRR6 NUP85-tdTomato cell (enlarged cell in (f)) incubated for 1 h with galactose. (h) Immunoblot of PGal1-APQ12-6His yeGFP-BRR6 NUP85-tdTomato cells. The pGal1 promoter was induced by the addition of galactose (t = 0). Samples were taken after the indicated times. Tub2 is a loading control. (i,j) Overexpression of APQ12 causes mislocalization of the ER protein Sec63-yeGFP (i) and Ole1-yeGFP (j). Cells with either the vector control or PGal1-APQ12-6His were incubated with galactose for the indicated time. (c,f,i and j) Scale bars: 5 µm.