Figure S2.
Phenotypic analysis of additional Ena mutants.(A) Graphs showing the expression levels of indicated actin nucleators throughout development (x axis; Ad, Adult). Units on the y axis are arbitrary. See Alyagor et al. (2018) for technical details. (B) Scheme depicting the genomic locus of ena. enaΔ151 is a CRISPR/Cas9-mediated one-nucleotide deletion located 151 bases downstream of the start site of isoforms B and F, enaLL03568 is a piggybac insertion located in the sixth coding exon, ena23 is a point mutation in the second coding exon causing an amino acid change of N379F, and ena210 is a point mutation in the seventh coding exon causing an amino acid change of A97V. Red and gray bars depict coding and noncoding exons, respectively, while lines depict introns. (C–E) Confocal z-projections of ena23 (C), ena210 (D), and enaLL03568 (E) adult γ-neuron MARCM neuroblast clones. (F) Growth index of WT (n = 10 medial lobe, n = 9 medial lobe) and enaΔ151 (n = 7) larval γ-neuron MARCM clones of both medial and dorsal lobes. The two groups were compared using two-tailed Student’s t test. Boxes encompass the values in between the first and third quartiles; whiskers are ±1.5 IQR; median (line), mean (blue triangle), and outliers (red diamond). (G and H) Confocal z-projections of WT (G) or enaΔ151 (H) larval γ-neuron MARCM clones. (I) Confocal single slice of the MB cell body region of brains containing an enaΔ151 γ-neuron MARCM neuroblast clone at 24 h APF and stained against Ena. I1 shows colabeling of the mutant clone (green) and anti-Ena (magenta). I2 is anti-Ena with the mutant clone depicted as a dotted line. (J) Confocal single slices of the MB cell body region depicting antibody staining of Ena in WT γ-neuron MARCM neuroblast clone at L3. J1 shows colabeling of Ena (magenta) and GFP. J2 shows antibody staining with the mutant clone demarcated with a white dashed line. Green is 71G10-Gal4–driven mCD8::GFP. Magenta is FasII in all panels except for I and J, where it is anti-Ena staining. Gray is 71G10-Gal4–driven mCD8::GFP in G and H, and.anti-Ena staining in I and J. Scale bars, 20 µm in all panels except in I and J, where they are 10 µm. Fhos, Formin homology 2 domain containing. A.U., arbitrary units; n.s., not significant.

Phenotypic analysis of additional Ena mutants. (A) Graphs showing the expression levels of indicated actin nucleators throughout development (x axis; Ad, Adult). Units on the y axis are arbitrary. See Alyagor et al. (2018) for technical details. (B) Scheme depicting the genomic locus of ena. enaΔ151 is a CRISPR/Cas9-mediated one-nucleotide deletion located 151 bases downstream of the start site of isoforms B and F, enaLL03568 is a piggybac insertion located in the sixth coding exon, ena23 is a point mutation in the second coding exon causing an amino acid change of N379F, and ena210 is a point mutation in the seventh coding exon causing an amino acid change of A97V. Red and gray bars depict coding and noncoding exons, respectively, while lines depict introns. (C–E) Confocal z-projections of ena23 (C), ena210 (D), and enaLL03568 (E) adult γ-neuron MARCM neuroblast clones. (F) Growth index of WT (n = 10 medial lobe, n = 9 medial lobe) and enaΔ151 (n = 7) larval γ-neuron MARCM clones of both medial and dorsal lobes. The two groups were compared using two-tailed Student’s t test. Boxes encompass the values in between the first and third quartiles; whiskers are ±1.5 IQR; median (line), mean (blue triangle), and outliers (red diamond). (G and H) Confocal z-projections of WT (G) or enaΔ151 (H) larval γ-neuron MARCM clones. (I) Confocal single slice of the MB cell body region of brains containing an enaΔ151 γ-neuron MARCM neuroblast clone at 24 h APF and stained against Ena. I1 shows colabeling of the mutant clone (green) and anti-Ena (magenta). I2 is anti-Ena with the mutant clone depicted as a dotted line. (J) Confocal single slices of the MB cell body region depicting antibody staining of Ena in WT γ-neuron MARCM neuroblast clone at L3. J1 shows colabeling of Ena (magenta) and GFP. J2 shows antibody staining with the mutant clone demarcated with a white dashed line. Green is 71G10-Gal4–driven mCD8::GFP. Magenta is FasII in all panels except for I and J, where it is anti-Ena staining. Gray is 71G10-Gal4–driven mCD8::GFP in G and H, and.anti-Ena staining in I and J. Scale bars, 20 µm in all panels except in I and J, where they are 10 µm. Fhos, Formin homology 2 domain containing. A.U., arbitrary units; n.s., not significant.

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