Extended Data Fig. 6: Comparison of the current protocol with the traditional FISH protocol combined with antibody staining in the liver and meningeal lymphatic and vascular system at the larval stage.

a, The liver after triple labelling of FISH-cp, 2F11, anti-GFP and with DAPI staining under the Tg(RasGFP) transgenic line at 5 dpf (3D stack of images; 104 µm, 26 slices, 4 µm each slice) using this protocol. b, The liver after triple labelling of FISH-cp, 2F11, anti-GFP and with DAPI staining under the Tg(RasGFP) transgenic line at 5 dpf (2D image) using this protocol. c, The liver after triple labelling of FISH-cp, 2F11, anti-GFP and with DAPI staining under the Tg(RasGFP) transgenic line at 5 dpf (3D stack of images; 108 µm, 27 slices, 4 µm each slice) using the traditional FISH protocol combined with antibody staining. d, The liver after triple labelling of FISH-cp, 2F11, anti-GFP and with DAPI staining under the Tg(RasGFP) transgenic line at 5 dpf (2D image) using the traditional FISH protocol combined with antibody staining. Note that the 2F11 and anti-GFP are hardly detectable with the traditional protocol. e, The meningeal lymphatic and vascular cells after triple labelling of the Dig-labelled mrc1a probe, anti-Kaede and anti-CFP in the Tg(lyve1b:Kaede; kdrl:CFP⁻NTR) transgenic line at 5 dpf using this protocol (3D stack of images; 200 µm, 50 slices, 4 µm each slice). f, The meningeal lymphatic and vascular cells after triple labelling of Dig-labelled mrc1a probe, anti-Kaede and anti-CFP in the Tg(lyve1b:Kaede; kdrl:CFP-NTR) transgenic line at 5 dpf using traditional FISH protocol combined with antibody staining (3D stack of images; 200 µm, 50 slices, 4 µm each slice). Note that the signals of FISH-mrc1a, anti-CFP and anti-Kaede are weak in the traditional protocol compared with this protocol. Scale bar, 100 µm.