FIGURE 3.

Top four panels, the eYFP tag on PLCβ2 does not affect its cellular localization. The upper left panel shows HEK293 cells expressing eYFP-PLCβ2 and the upper right panel shows cells expressing untagged PLCβ2 as detected by immunostaining with Alexa647-anti-PLCβ2. The middle panels show the low amount of endogenous PLCβ2 in HEK293 cells under basal and stimulated conditions (5 μm carbachol) in fixed cells as detected using Alex647-anti-PLCβ2. Bottom, changes in the localization of eCFP-PLCδ1 (top panels) and eYFP-PLCβ2 (bottom panels) expressed in HEK293 cells with 5 μm carbachol stimulation. Although some movement of eCFP-PLCδ1 toward the plasma membrane occurs in a time-dependent manner, eYFP-PLCβ2 remains cytosolic. All images were viewed through a 60× objective (see “Materials and Methods”).