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. 2019 May 9;15(5):e1008153. doi: 10.1371/journal.pgen.1008153

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© 2019 Shyu et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 3 — (A) Expression patterns (green) of VT30604-GAL4 (α′β′ neurons), VT44966-GAL4 (γ neurons), and C739-GAL4 (αβ neurons). The brain structures were counterstained with a DLG antibody (magenta). The scale bars represent 50 μm. (B) VT30604-GAL4, VT44966-GAL4, and C739-GAL4 were used to drive UAS-inx5^RNAi expression in MB α′β′, γ, or αβ neurons, respectively. The 3-hour ARM values for these flies, including controls, are shown. Each value represents the mean ± SEM (n = 8). *, p < 0.05; one-way analysis of variance (ANOVA) followed by Tukey’s test. The genotypes were as follows: (1) +/+; +/+, (2) +/+; UAS-inx5^RNAi(v6950)/+, (3) +/+; VT30604-GAL4/+, (4) +/+; VT30604-GAL4/UAS-inx5^RNAi(v6950), (5) +/+; VT44966-GAL4/+, (6) +/+; VT44966-GAL4/UAS-inx5^RNAi(v6950), (7) C739-GAL4/+; +/+, and (8) C739-GAL4/+; +/UAS-inx5^RNAi(v6950). (C) The expression pattern of C739-GAL4-driven GFP expression in the central brain. The brain structures were counterstained with a DLG antibody (magenta). The scale bar represents 50 μm. (D) C739-GAL4-driven GFP expression in αβ neurons was reduced using the MB-GAL80 transgene. The brain structures were counterstained with a DLG antibody (magenta). The scale bar represents 50 μm. (E) ARM defect in the C739;inx5^RNAi(v6950)/+ flies was rescued by removing the expression of GAL4 in αβ neurons using the MB-GAL80 transgene. Each value represents the mean ± SEM (n = 8) *, p < 0.05; ANOVA followed by Tukey’s test. The genotypes were as follows: (1) +/+; +/+, (2) MB-GAL80/+; +/+, (3) C739-GAL4/+; UAS-inx5^RNAi(v6950)/+, and (4) C739-GAL4/MB-GAL80; +/UAS-inx5^RNAi(v6950). (F) Expression pattern of VT49246-GAL4 (αβ neurons). The brain structures were counterstained with a DLG antibody (magenta). The scale bars represent 50 μm. (G) VT49246-GAL4 was used to drive UAS-inx5^RNAi(v6950) to confirm the requirement for INX5 in MB αβ neurons for 3-hour ARM formation. Each value represents the mean ± SEM (n = 13–14). *, p < 0.05; ANOVA followed by Tukey’s test. The genotypes were as follows: (1) +/+; +/+, (2) +/+; VT49246-GAL4/+, (3) +/+; UAS-inx5^RNAi(v6950)/+, and (4) +/+; VT49246-GAL4/UAS-inx5^RNAi(v6950).