1. 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.

2. 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.

3. 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.

4. 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.