Fig. 1: Recruitment of FKBP-BcPI-PLC3A to the OMM induces localized DAG production and rapid fragmentation of the mitochondrial network. | Nature Communications

Fig. 1: Recruitment of FKBP-BcPI-PLC3A to the OMM induces localized DAG production and rapid fragmentation of the mitochondrial network.

From: Acute diacylglycerol production activates critical membrane-shaping proteins leading to mitochondrial tubulation and fission

Fig. 1

a, b Representative images of HEK293A cells (5 μm scale bar) showing localization of the OMM-targeted FRB recruiter (OMM-FRB-ECFP, magenta) and a high-affinity DAG-binding probe (NES-mEGFP-MmPKDC1a,b, green) in response to rapamycin-induced (100 nM) recruitment of either the catalytically active (a, mRFP-FKBP-BcPI-PLC3A) or inactive (b, mRFP-FKBP-BcPI-PLCDEAD) enzyme variants (gray) to the cytosolic membrane leaflet of the mitochondria. The composite images present an overlay of the OMM-FRB together with the DAG biosensor. c Kinetics of DAG production within the OMM of HEK293A cells following recruitment of iRFP-FKBP-BcPI-PLC3A (green trace) or iRFP-FKBP-BcPI-PLCDEAD (gray trace) to the cytosolic membrane leaflet of the mitochondria, as measured using the OMM-DAGBRET biosensor (AKAPTM-mVenus-T2A-sLuc-NES-MmPKDC1a,b). Population-level measurements in HEK293A defining the OMM-FRB:FKBP dimerization kinetics (red trace) for the FKBP-BcPI-PLCDEAD scaffold upon rapamycin-induced (100 nM) recruitment to the OMM, as measured using the OMM-FRB:FKBPBRET biosensor (AKAPTM-FRB-mVenus-T2A-sLuc-FKBP-BcPI-PLCDEAD; red trace). BRET measurements are presented as mean values ± SEM from three independent experiments carried out using triplicate wells.

Back to article page