The role of Wash in terminal cell development. (A) Terminal cell expressing CD4::mIFP and Wash::GFP under btl-gal4. The region marked in A is shown in A’–A’’’ at higher magnification over eight time points. (B–F) Cells expressing PH::GFP under btl-gal4; control (B) and wash knockdowns (C and D). (E) Proportion of cells with the indicated phenotypes. Number of cells analyzed: control, n = 29; wash-IR, n = 46. Significance was assessed with χ² test; ****, P > 0.0001. (F) Distance between the tip of the tube to the tip of the cell in cells with a normal, ventrally guided tube. Number of cells analyzed: control, n = 28; wash-IR,n = 35. Significance was assessed with a Mann-Whitney U test. (G–J)shrb^G5 heterozygous embryos expressing CD4::mIFP under btl-gal4; shrb^G5 heterozygote as control (G) and shrb^G5 heterozygote expressing wash-IR (H and I). (H) Cell with a misguided tube. (I and I’) Cell lacking a tube is shown at two time points. (J) Proportion of cells with the indicated phenotypes. Number of cells analyzed: shrb^G5 heterozygotes (controls), n = 30; shrb^G5 heterozygotes expressing wash-IR, n = 34. Significance was assessed with χ² test; ****, P > 0.0001. (K–M) Cells expressing CD4::mIFP and Wash::GFP under btl-gal4. (K and L) Control (K and K’), cell also expressing Vha100^R755A (V100^R755A; L and L’). Arrowheads point to CD4 vesicles. (M) Fluorescence intensity of Wash::GFP at CD4 vesicles normalized over the mean signal of Wash::GFP in the cytoplasm. Number of cells analyzed: control, n = 11; Vha100^R755A, n = 17. Significance was assessed with a Mann-Whitney U test. Box plots in F and M represent median, IQR, and IQR*1.5 below and above the IQR. Ventral is down.