Fig 4.
PikA disrupts the distribution of PI3P in cells and colocalizes with PI5P. (A) Distribution of the PI3P probe GFP-2xFYVEHrs in cells expressing and mCherry-PikA, mCherry-PikAH171A, or mCherry-PikAD194A. HeLa cells transfected to express the probe and mCherry-PikA or its mutants for 14 h were used to acquire images. Nuclei were stained with Hoechst (left). Bar, 5 µm. Quantitation of cells with an altered distribution of the PI3P probe. Results (mean ± s.e.) shown were from three independent experiments done in triplicate, at least 100 cells were counted for each sample (right). (B) Distribution of the PI5P probe Dok1-GFP in cells expressing mCherry-tagged PikA or its enzymatically inactive mutants. Samples were prepared as described in A (left). Bar, 5 µm. Quantitation of the co-localization between the PI5P probe and PikA or its mutants. Results (mean ± s.e.) shown were from three independent experiments done in triplicate, at least 100 cells were counted for each sample (right). ***, P < 0.001.