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Figure 4


Fig. 4. The effect of PIP5K and 5-phosphatase on the location of PLCβ1a. (A,B) HeLa cells coexpressing GFP-PIP5K (imaged separately in A) and RFP-PLCβ1a (imaged separately in B). (C) The fluorescence intensity ratio of PLC to PIP5K was calculated for cells in which RFP-PLCβ1a does [TL (translocation)] and does not (no TL) dissociate from the plasma membrane upon stimulation with 100 µM histamine. A two-tailed Student's t-test (confidence level of 95%) indicates that the cells in which PLCβ1a does not translocate to the cytosol express significantly more PIP5K than those in which PLCβ1a diffuses into the cytosol. Error bars depict standard error. (D,E,G,H) HeLa cells coexpressing untagged 5-phosphatase IV, YFP-PLCβ1a (D,G) and CFP-PH{delta}1 (E,H). (F) The ratio between plasma membrane fluorescence and cytosolic fluorescence of YFP-PLCβ1a was calculated for cells expressing PLCβ1a alone (PLC), in the presence of PIP5K (PLC+PIP5K) and in the presence of 5-phosphatase (PLC+5ptase). A two-tailed Student's t-test (confidence level of 95%) indicates that significantly more PLCβ1a localizes at the plasma membrane in the presence of PIP5K, and that significantly less localizes at the plasma membrane in the presence of 5-phosphatase. Error bars depict standard error.





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