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Fig. 1. vari/CG9326 encodes a MAGUK required for tracheal tube size control. (A,B,E,F,H,I) Compared to wild type, vari mutant trachea are long (B,F) and are missing lumenal segments of the ganglionic branches (bracket in I). (C,D) RNAi knockdown of CG9326 phenocopies vari mutations. (G,J) Ubiquitous expression of the short vari isoform rescues vari null mutant phenotypes. (K) The vari/CG9326 genomic locus, transcript organization, protein domains and mutations. , alternative splicing generates either a 35 or 14 amino acid linker between the PDZ and SH3 domains. Note the PDZ-SH3 linker in PALS2, a mouse homolog of Vari, is also alternatively spliced to produce linkers of different length (Kamberov et al., 2000); ^*, splicing at a GC splice donor site was confirmed by sequencing seven independent cDNAs and the genomic DNA of multiple wild-type and lab stocks. (L-N) Varicose (green) co-localizes with the SJ marker Nrx (red) in wild-type (vari^F033/+) surface epithelia (L), but is absent in genetically null mutants vari^F033 (M) and vari^R3 (N). Left and right panels show the same image, but left panels show guinea pig anti-Vari (green channel), whereas right panels show Vari and Nrx (composite red/green). Trachea visualized in A,B,F-J with monoclonal 2A12, in C,D with btl::gal4 UAS GFP. Genotype in G,J is vari^F033 da::gal4/vari^F033 UAS vari short. (A-J,L-N) Stage 16 animals. Dashed lines indicate location of basal cell surfaces; L and M are cropped from the same image of adjacent heterozygous and homozygous animals. Scale bars: in B, 25 µm for A,B; in J, 50 µm for C-J; in N, 1.5 µm for L-N. ORF, open reading frame; UTR, untranslated region; WT, wild type.

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