The localisation of the PIP<sub>2</sub> sensor, Tubby-YFP, in HEK293 cells before and after the rapamycin-induced recruitment of PJ or PJ-DEAD to the PM.
<p>The rapamycin (Rap)-induced dimerization of PJ and PJ-DEAD with PM-localised LYN<sub>11</sub>-FRB was investigated in transiently transfected HEK293 cells. Tubby-YFP localises to the PM in the presence of a sufficient PIP<sub>2</sub> concentration. <b>A</b> and <b>B</b>. Top panel: Before rapamycin addition. Lower panels: Increasing time after addition of rapamycin showing the increased PM signal of PJ and PJ-DEAD. Upon recruitment of PJ to the PM, Tubby-YFP redistributes from the PM to the cytosol (<b>A</b>). Upon recruitment of PJ-DEAD to the PM, Tubby-YFP remains PM-localised (<b>B</b>). Scale bar indicates 15 μm. <b>C–F</b>. Quantified line (<b>C</b> and <b>E</b>) and box plots (<b>D</b> and <b>F</b>) from the indicated red lines and boxed areas in <b>A</b> and <b>B</b> (located in the top right hand merged panels), highlighting translocation of Tubby-YFP to the cytosol when PJ (<b>C</b> and <b>D</b>) but not PJ-DEAD (<b>E</b> and <b>F</b>) is recruited to the PM. Red arrows indicate rapamycin addition.</p
