Figure 5.
Figure 5. PI4KIIIα and PtdIns4P production. (A) Western blot analysis of the four PI 4-kinase isoforms in lysates from control (Ctrl) and PI4KIIIα KO MEFs. (B and C) Quantification of PtdIns4P and PtdIns(4,5)P2 (shown as a percentage of total inositol phospholipids, n = 3) in control and PI4KIIIα KO MEFs, respectively. *, P < 0.0001. (D) Imaging of the plasma membrane PtdIns4P pool in control and PI4KIIIα KO MEFs using GFP-PHOsh2. (E) Quantification of the plasma membrane/cytosol (PM/cyt) GFP-PHOsh2 fluorescence ratio in D based on line scans (control, n = 29; KO, n = 30; *, P < 0.0001). (F) Imaging of the Golgi complex PtdIns4P pool in control and PI4KIIIα KO MEFs using GFP-GOLPH3. (G) Imaging of Golgi complex morphology in control and PI4KIIIα KO MEFs using anti-GM130 immunofluorescence. Error bars indicate standard deviations. Bars, 20 µm.

PI4KIIIα and PtdIns4P production. (A) Western blot analysis of the four PI 4-kinase isoforms in lysates from control (Ctrl) and PI4KIIIα KO MEFs. (B and C) Quantification of PtdIns4P and PtdIns(4,5)P2 (shown as a percentage of total inositol phospholipids, n = 3) in control and PI4KIIIα KO MEFs, respectively. *, P < 0.0001. (D) Imaging of the plasma membrane PtdIns4P pool in control and PI4KIIIα KO MEFs using GFP-PHOsh2. (E) Quantification of the plasma membrane/cytosol (PM/cyt) GFP-PHOsh2 fluorescence ratio in D based on line scans (control, n = 29; KO, n = 30; *, P < 0.0001). (F) Imaging of the Golgi complex PtdIns4P pool in control and PI4KIIIα KO MEFs using GFP-GOLPH3. (G) Imaging of Golgi complex morphology in control and PI4KIIIα KO MEFs using anti-GM130 immunofluorescence. Error bars indicate standard deviations. Bars, 20 µm.

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