Figure 6.
PI4KII is required for accumulation of PI4P on membranes of mature SGs.(A, C, E, G, and H) Leica point-scanning confocal images of live late L3 salivary gland cells processed with Leica Lightning algorithm. (A–A′′) Control cells expressing CD63-GFP (green) and mCh-PI4KII (red). Contacts formed by mCh-PI4KII tubules and CD63-GFP–positive SG membranes are marked by magenta arrowheads. (B) Colocalization analysis of CD63-GFP and mCh-PI4KII. Pearson’s correlation coefficient = −0.053 ± 0.025, M(CD63) (Mander’s overlap coefficient of CD63-GFP) = 0.042 ± 0.0075, M(PI4KII) = 0.36 ± 0.11. Error bars indicate standard deviation. (C–C′′) Control cells expressing CD63-GFP (green) and mCh-2xP4M (red). Strong colocalization of CD63-GFP and mCh-2xP4M (yellow) is marked by magenta arrowheads. (D) Colocalization analysis of CD63-GFP and mCh-2xP4M. Pearson’s = 0.41 ± 0.067, M(CD63) = 0.51 ± 0.061, M(2xP4M) = 0.43 ± 0.084. Error bars indicate standard deviation. (E–E′′) Control cells expressing GFP-PI4KII (green) and mCh-2xP4M (red). Colocalization of GFP-PI4KII and mCh-2xP4M on SG membranes is marked by magenta arrowheads. (F) Colocalization analysis of GFP-PI4KII and mCh-2xP4M. Pearson’s = 0.32 ± 0.085, M(PI4KII) = 0.90 ± 0.036, M(2xP4M) = 0.36 ± 0.050; ****, P < 0.0001. Error bars indicate standard deviation. (G and H) Cells expressing Sgs3-GFP (green) and mCh-2xP4M (red) in control (G) or PI4KIIΔ mutant (H). SGs coated by mCh-2xP4M are marked by magenta arrowheads. mCh-2xP4M vesicle lacking Sgs3-GFP is marked by yellow arrowhead. (I) Percentage of SGs coated by mCh-2xP4M. n = one cell each from three independent salivary glands; ***, P < 0.001. Error bars indicate standard deviation.

PI4KII is required for accumulation of PI4P on membranes of mature SGs. (A, C, E, G, and H) Leica point-scanning confocal images of live late L3 salivary gland cells processed with Leica Lightning algorithm. (A–A′′) Control cells expressing CD63-GFP (green) and mCh-PI4KII (red). Contacts formed by mCh-PI4KII tubules and CD63-GFP–positive SG membranes are marked by magenta arrowheads. (B) Colocalization analysis of CD63-GFP and mCh-PI4KII. Pearson’s correlation coefficient = −0.053 ± 0.025, M(CD63) (Mander’s overlap coefficient of CD63-GFP) = 0.042 ± 0.0075, M(PI4KII) = 0.36 ± 0.11. Error bars indicate standard deviation. (C–C′′) Control cells expressing CD63-GFP (green) and mCh-2xP4M (red). Strong colocalization of CD63-GFP and mCh-2xP4M (yellow) is marked by magenta arrowheads. (D) Colocalization analysis of CD63-GFP and mCh-2xP4M. Pearson’s = 0.41 ± 0.067, M(CD63) = 0.51 ± 0.061, M(2xP4M) = 0.43 ± 0.084. Error bars indicate standard deviation. (E–E′′) Control cells expressing GFP-PI4KII (green) and mCh-2xP4M (red). Colocalization of GFP-PI4KII and mCh-2xP4M on SG membranes is marked by magenta arrowheads. (F) Colocalization analysis of GFP-PI4KII and mCh-2xP4M. Pearson’s = 0.32 ± 0.085, M(PI4KII) = 0.90 ± 0.036, M(2xP4M) = 0.36 ± 0.050; ****, P < 0.0001. Error bars indicate standard deviation. (G and H) Cells expressing Sgs3-GFP (green) and mCh-2xP4M (red) in control (G) or PI4KIIΔ mutant (H). SGs coated by mCh-2xP4M are marked by magenta arrowheads. mCh-2xP4M vesicle lacking Sgs3-GFP is marked by yellow arrowhead. (I) Percentage of SGs coated by mCh-2xP4M. n = one cell each from three independent salivary glands; ***, P < 0.001. Error bars indicate standard deviation.

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