Figure 3. Plasma membrane localized... | Rockefeller University Press

Figure 3.

$Figure 3. Plasma membrane localized PI(4,5)P2 depends on Mss4. (A) The GFP-PHPlcδ-PHPlcδ-GFP reporter localizes to the PM, dependent on Mss4. Images (top, DIC; bottom, central z-section GFP fluorescence) of representative mss4 cells grown as indicated. (B) Expression level of the PI(4,5)P2 reporter is not affected by reduction of Mss4 level. Immunoblot showing the levels of reporter (α-GFP) and loading control (α-Cdc11), strains and conditions as in A. (C) Quantitation of cells with PM localized PI(4,5)P2. Average percentage of cells with PM localized reporter (ni.ex. = 2–3; 4 determinations, n = 200 cells each) indicated. (D) PI(4,5)P2 levels are reduced in mss4 mutant. PI(4,5)P2 levels from 100,000-g fraction determined in indicated strains using an anti-PI(4,5)P2 mAb (ni.ex. = 2; 4 determinations each).$

Plasma membrane localized PI(4,5)P₂ depends on Mss4. (A) The GFP-PH^Plcδ-PH^Plcδ-GFP reporter localizes to the PM, dependent on Mss4. Images (top, DIC; bottom, central z-section GFP fluorescence) of representative mss4 cells grown as indicated. (B) Expression level of the PI(4,5)P₂ reporter is not affected by reduction of Mss4 level. Immunoblot showing the levels of reporter (α-GFP) and loading control (α-Cdc11), strains and conditions as in A. (C) Quantitation of cells with PM localized PI(4,5)P₂. Average percentage of cells with PM localized reporter (n_i.ex. = 2–3; 4 determinations, n = 200 cells each) indicated. (D) PI(4,5)P₂ levels are reduced in mss4 mutant. PI(4,5)P₂ levels from 100,000-g fraction determined in indicated strains using an anti-PI(4,5)P₂ mAb (n_i.ex. = 2; 4 determinations each).