Fig. 2

LRR/CT-dependent Adgra2 intracellular trafficking. (A) Schematic representation of Adgra2, Adgra2^?LRR1, Adgra2^?LRR2, Adgra2^?LRR4, Adgra2^?LRR, Adgra2^?Ig-like, Adgra2^?HRM, Adgra2^?GAIN, Adgra2^?LRR/CT, Adgra2^{?LRR/CT/Ig-like} and Adgra2^{?LRR/CT/Ig-Like/HRM} domain organization. See Fig. 1A for schematic labels. (B) Single-plane direct fluorescence confocal scans of non-permeabilized HEK293T cells 48?h after transfection with the indicated adgra2 variants together with the GPI-RFP membrane marker or the mCherry-SEC61? ER marker. Cells were additionally transfected with reck and Wnt7a (mouse gene) expression constructs. Nuclei were counterstained with Hoechst. Scale bar: 10??m. (C) Colocalization assessment of Adgra2 and its variants with the membrane marker GPI-RFP (red dots) or the ER marker mCherry-SEC61? (blue dots) using the Pearson correlation coefficient. Error bars represent median±interquartile range. (D) Quantification of neurog1:EGFP⁺ DRG at 72?hpf (red dots) and hindbrain CtAs at 60?hpf (blue dots) in WT and adgra2 morphant larvae and embryos injected at the one-cell stage with 100?pg RNA encoding Adgra2 or Adgra2 variants. Error bars represent median±interquartile range (***P<0.001; ****P<0.0001; Kruskal?Wallis test).