Fig. 4.

Increased Yap1801/Yap1802 increases rates of cargo endocytosis. (A,B) Airyscan confocal microscopy imaging of wild-type cells co-expressing Mup1–GFP (A) or Can1–GFP (B) and Yap1801–mCherry or Yap1802–mCherry, expressed under control of the copper-inducible CUP1 promoter. The mean±s.d. percentages of cells exhibiting exclusive plasma membrane localisation for each condition (>75 cells per conditions) are shown in red text. (C) Localisation of Mup1–GFP in wild-type, mig1Δ, and mig1Δ mig2Δ cells at mid- and late-log phase. (D) Quantification of the experiment shown in C, with 35–120 mid-log phase cells quantified from each condition. Mean±s.d. n=3. (E) Immunoblot analysis of Mup1–GFP expressed in wild-type and mig1Δ mig2Δ cells. GAPDH was used as a loading control. (F) Wild-type and mig1Δ mig2Δ cells expressing Can1–GFP, Ste3–GFP and Fur4–mNG were imaged at mid- and late-log phase. Surface-localised signal is indicated with white arrows. (G) Immunoblot analysis of vacuolar GFP from lysates generated from wild-type and yap1801Δ yap1802Δ cells expressing Mup1-GFP. GAPDH was used as a loading control. (H) Mup1–GFP was expressed in wild-type cells labelled with Sec7–mCherry and in unlabelled yap1801Δ yap1802Δ cells, mixed in a 1:1 ratio and co-cultured for 2 h, followed by 90 additional minutes in glucose (left) or raffinose (right) media prior to fluorescence microscopy. Surface-localised signal is indicated with yellow arrowheads. (I) Quadruple-null mig1Δ mig2Δ yap1801Δ yap1802Δ cells expressing Mup1–GFP, Can1–GFP, Ste3–GFP or Fur4–mNG were imaged at mid-log phase. Scale bars: 5 µm.