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. 2018 Feb 27;7:e33555. doi: 10.7554/eLife.33555

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© 2018, Koe et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 6. — (A) In vivo BiFC assay between Vib and Sqh. NYFP-Myc-Vib and CYFP-HA-Sqh were co-expressed in neuroblasts by insc-Gal4, stained with HA (red), Myc (red) and DAPI (grey), and detected for YFP fluorescence (green). Controls were NYFP-Myc-Vib with CYFP-HA control and CYFP-HA-Sqh with NYFP-Myc Control. Scale bars, 5 µm. (B) Schematic representation of proximity ligation assay performed on S2 cells (refer to Materials and methods). (C) In situ PLA assay between Flag-Vib and Myc-Sqh in S2 cells. S2 cells transfected with the indicated plasmids were stained with Flag (green), Myc (blue) and DAPI (grey) and detected for PLA signal (red). Cell outline was shown by differential interference contrast (DIC) images. Scale bar, 5 µm. (D) Graph showing the percentage of S2 cells that expressed no PLA signal, weak (≤10 foci), moderate (11–30 foci) and strong (>30 foci) PLA signals for (C). Quantification for the average number of PLA foci per cell in (C).

Figure 6—figure supplement 1. — (A) In vivo BiFC assay between Vib and Sqh. NYFP-Myc-Vib and CYFP-HA-Sqh were co-expressed in neuroblasts by insc-Gal4, stained with HA (red), Myc (red) and DAPI (grey), and detected for YFP fluorescence (green). Controls were NYFP-Myc-Vib with CYFP-HA control and CYFP-HA-Sqh with NYFP-Myc Control. Scale bars, 5 µm. (B) Schematic representation of proximity ligation assay performed on S2 cells (refer to Materials and methods). (C) In situ PLA assay between Flag-Vib and Myc-Sqh in S2 cells. S2 cells transfected with the indicated plasmids were stained with Flag (green), Myc (blue) and DAPI (grey) and detected for PLA signal (red). Cell outline was shown by differential interference contrast (DIC) images. Scale bar, 5 µm. (D) Graph showing the percentage of S2 cells that expressed no PLA signal, weak (≤10 foci), moderate (11–30 foci) and strong (>30 foci) PLA signals for (C). Quantification for the average number of PLA foci per cell in (C).