A B C D E F Supplementary Figure S1 Deng et al. MS1096

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A B C D E F Supplementary Figure S1 Deng et al. MS1096 MS1096 >HpoVC MS1096 >HpoVN MS1096 >HpoVC HpoVN MS1096 >SavVN MS1096 >HpoVC SavVN A B C D E F Adult fly wing analysis shows the modification of BiFC to protein has no obvious influence to their function in growth inhibition. Expression of the transgenes was driven by a wing-specific Gal4 driver, MS1096.

A B C D E F G Supplementary Figure S2 Deng et al. HpoVC HpoVN HpoT189AVC HpoT189AVN HpoT195AVC HpoT195AVN HpoT189AT195AVC HpoT189AT195AVN HpoT189EVC HpoT189EVN HpoT195EVC HpoT195EVN HpoT189ET195EVC HpoT189ET195EVN Mutation of Thr189 and Thr195 auto-phosphorylation sites to Ala or Glu does not influence the Hpo BiFC signal. The Hpo constructs indicated above were expressed in fly S2 cells for BiFC analysis.

B C Supplementary Figure S3 Deng et al. D E F C5>Flag-Hpo WT G H I E-Cad Merge D E F C5>Flag-Hpo WT Hpo-Hpo BIFC B C HA-Hpo G H I J K L M N DAPI Detection of Hpo and Hpo dimer in developing wing tissues. (A-C) Flag-tagged Hpo (green) was stained in C5-Gal4/UAS-Flag-hpo third-instar larval wing discs. (D-F) Endogenous Hpo (green) was detected in wild-type larval wing discs. DE-cadherin (red) in (A-F) was a marker for the plasma membrane. (G-N) While endogenous Hpo and HA-Hpo (red) are usually distributed throughout the cell, Hpo dimer (green) shows a more membrane-associated pattern in many cells. The BiFC signal was generated in en-Gal4/UAS-hpoVC hpoVN larval wing discs. DE-cadherin and the nucleus were detected in blue in (H) and (L), respectively. Arrows identify some specific examples.

A B C Supplementary Figure S4 Deng et al. HA-mer HA-mer mer RNAi Control RNAi HA-ex HA-ex ex RNAi Control RNAi Myc-kibra Myc-kibra kibra RNAi Control RNAi WB: a-HA WB: a-Ex WB: a-Myc A B C Drosophila S2 cells were transfected with HA-mer, HA-ex, or Myc-kibra. Specific RNAi treatment was done in order to abolish or reduce expression of these genes. The results show that the RNAi treatment can be effective in decreasing Mer, Ex, and Kibra expression levels in S2 cells. mCherry RNAi was used as control RNAi.