Fig. 1

Fig. 1

a Cartoon representation of GFP/YFP SPLICS_S/L-P2A probes. The cartoon also indicates the occurrence of reconstitution of the two spectral variants through association with the invariant strand β₁₁. b Schematic representation of the targeted GFP/YFP SPLICS_S/L-P2A chimeras. GFP/YFP protein was split in two non-fluorescent portions, G(Y)FP_1–10 and β11 fragment that are targeted to outer face of organelles of interest. The complementation of targeted split-GFP at membrane contact sites was also shown in the cartoon. c Creation of a single vector for the equimolar expression of the targeted fragments through the insertion of the P2A peptide. d–g Schematic representations of the SPLICS_S/L-P2A vectors. The β11_S/L coding sequence is cloned upstream the viral P2A peptide sequence, while the second cassette is occupied by the G(Y)FP_1–10. d SPLICS_S/L-P2A^ER–MT expression plasmids (ER_S/L-β11 and OMM-GFP_1–10). e SPLICS_S/L-P2A^ER–PM expression plasmids (ER_S/L-β11 and PM-YFP_1–10). f SPLICS_S/L-P2A^PO–MT expression plasmids (PO_S/L-β11 and OMM-GFP_1–10). g SPLICS_S/L-P2A^PO–ER expression plasmids (PO_S/L-β11 and ER-YFP_1–10).