Figure 1.

Mts physically interacts with Vtd. (a) Confirmation of positive interaction between C-terminal part of Mts (MtsC67) and Vtd fragment by yeast two-hybrid assay. Vtd alone shows no lacZ-positive blue colonies on the media plate lacking leucine and tryptophan (-Leu/-Trp/X-α-Gal plate) or the media deficient in adenine, histidine, leucine and tryptophan (-Ade/-His/-Leu/-Trp/X-α-Gal/Aureobasidin A(AbA)). Direct interaction between MtsC67 and Vtd results in the formation of blue colonies on both selection media plates. (b) GST pull-down assay with MBP-Mts^WT and GST-Vtd^WT. Mts and Vtd show direct interaction in vitro. (c) Co-immunoprecipitation assay using S2 cell extract. FLAG-tagged Mts^WT forms a protein complex with V5-tagged Vtd^WT in S2 cell. (d) Co-immunoprecipitation assay using embryo extract. Embryos are vtd mutants rescued by wild-type Vtd-Myc (w*; vtd^ex3, tub::vtd^WT-myc₁₀). Myc-tagged Vtd^WT co-immunoprecipitates with endogenous Mts^WT in embryos. (b–d) Full-length blots are presented in Supplementary Fig. 6.