FIGURE

Fig. 3

ID

ZDB-FIG-221118-178

Publication

Fasano et al., 2022 - Dominant ARF3 variants disrupt Golgi integrity and cause a neurodevelopmental disorder recapitulated in zebrafish

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Fig. 3

a Maximum intensity confocal z-projections showing immunostaining against Golgin-97 (trans-Golgi marker) (green) performed in fixed COS-1 cells transiently transfected with mCherry-tagged ARF3^WT, ARF3^T31N, and ARF3^Q71L (DN and CA variants, respectively) or mutants identified (magenta) for 48 h. Composite colocalization images are shown in the right panels with nuclei (DAPI staining) in blue. The images are representative of three independent experiments. Scale bars = 2 µm (high magnification) and 10 µm (all the other images). b Golgi means intensity and area define distinct Golgi morphotypes. Dot plots of mean intensity (MI) and area of Golgi in cells transiently transfected with mCherry-tagged ARF3^WT, or ARF3^Q71L and ARF3^T31N mutants (up, middle, bottom panels) are shown. Golgi MI and area (% of the whole cell) of cells were measured based on Golgin-97 staining. Whole-cell area was determined using the area covered by mCherry fluorescence as a mask. Representative 3D rendering images of the observed Golgi staining are shown. Cell populations located in different gates are characterized by distinct Golgi morphologies: Compact: A < 2.6 and MI > 1.5 (green gate); expanded Golgi: 2.7 < A < 12 and MI > 0.8, (purple gate); partially dispersed Golgi: 2.7 < A < 12 and MI < 0.8 (pink gate); totally dispersed Golgi: A > 12 and MI < 0.8 (bordeaux gate). c Incidence of trans-Golgi morphotypes. The bar graph represents the percentage of cells showing compact, expanded, partially or fully dispersed distribution (part.disp and ful.disp.) of Golgi in mCherry-tagged ARF3 transfected cells, based on the classification described above in (b). No. of cells = 26 (WT); 22 (Q71L); 29 (T31N); 20 (K127E); 20 (L12V/D67V); 21 (P47S); 28 (D93N) and 27 (T32N). Data are expressed as mean ± SEM of three independent experiments. Two-sided Chi-square’s test in a 2 × 2 contingency table (WT vs. all mutants, compact vs. all phenotypes ****p < 0.0001) is used to assess statistical significance. Arb.units = arbitrary units. Source data are provided as a Source Data file.

Expression Data

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Antibody Labeling

Phenotype Data

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Acknowledgments

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