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. 2022 May 17;221(7):e202110167. doi: 10.1083/jcb.202110167

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Figure S5. — Validation of expression of mutant NRG3 constructs and of goat polyclonal anti-NRG3 antibody used in this study. (A) Representative images showing expression of V5-tagged NRG3^Q360^*, hFB-NRG3, ICD^N-TM^N, ICD^N-TM^C, ICD^C-TM^C, and ICD^C-TM^N in transfected HEK293 cells. Cells were fixed and stained with anti-V5; DAPI nuclear stain was also included. (B) Still frames of live-cell imaging experiments in HEK293 cells co-transfected with GFP-NRG3 and either TM^N-mCherry or TM^C-mCherry. Note that TM^N-mCherry signals overlap extensively with GFP-NRG3 including in the cell periphery (white arrowheads) whereas TM^C-mCherry distribution is mostly non-overlapping with GFP-NRG3 (white/blue arrowheads). (C and D) Goat anti-NRG3 antibody labeling specificity was verified in sagittal sections from adult WT (C) and NRG3 knockout (D) mouse brains. Note the absence of mossy fiber staining in CA3 stratum lucidum (sl) in the KO section. Scale bars: A and B, 10 µm; C and D, 100 µm.