Fig. 6

Expression of the SPLICS_S probe in living zebrafish embryos. a Schematic depiction of the bidirectional construct that allows detection of DsRed and SPLICS_S-P2A in a Gal4-dependent manner. The 2A peptide guarantees the generation of an equimolar amount of the two spGFP fragments. b Experimental setting used to image ER–mitochondria contacts in zebrafish embryos: a schematic drawing of RB neurons is shown on the right. c Representative image of a 1 dpf s1102t:GAL4 embryo injected with the pT2-DsRed-UAS-SPLICS_S-P2A construct. d Live imaging of short ER–mitochondria contacts in RB neurons of s1102t:GAL4 zebrafish embryos. The picture is the merge of several planes. The 3D rendering of the z-stack is shown on the right. Scale bar: 15 µm. e Quantification of SPLICS_S contacts in the cell body of RB neurons by 3D rendering of complete z-stacks. Mean ± SEM: 22 ± 1, n = 28 cells from 11 fish. f Live imaging of SPLICS_S contacts in the axons of RB neurons. The picture is the merge of several planes. The 3D rendering of the complete z-stack is shown on the right. Scale bar: 15 µm. g Quantification of the density of SPLICS_S contacts in the cell body and the axons of RB neurons. Mean ± SEM: RB soma: 0.17 ± 0.01, n = 28 cells from 11 fish; RB axon: 0.14 ± 0.01, n = 20 cells from 6 fish. Data shown are the result of two independent experiments.