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Figure 1 | Journal of Negative Results in BioMedicine

Figure 1

From: Biological constraints limit the use of rapamycin-inducible FKBP12-Inp54p for depleting PIP2 in dorsal root ganglia neurons

Figure 1

Inp54p translocates to plasma membrane in HEK293 cells, reducing PIP 2 biosensor levels following rapamycin treatment. A-F) HEK293 cells were transfected with plasmids encoding FRBPLF-CFP, Venus-FKBP12-Inp54p and PLC∂1PH-RFP, a PIP2 biosensor. One day later, cells were imaged by live-cell confocal microscopy. Before rapamycin treatment, (A) FRBPLF-CFP, (B) Venus-FKBP12-Inp54p, and (C) PLC∂1PH-RFP were properly localized to the (A,C) membrane or (B) cytoplasm. While localization of FRBPLF-CFP remains constant, rapamycin (1 μM) induced rapid (<3 s) translocation of (E) Venus-FKBP12-Inp54p to the plasma membrane and (F) reduced PLC∂1PH-RFP levels at the membrane, indicative of PIP2 hydrolysis. G) Quantification of translocation in HEK293 cells. **P < 0.005 compared to pre-rapamycin condition, n = 20 cells per condition. Scale bar, 10 μm.

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