Quantiﬁcation of Apoptotic Indexes in Mutant Clones
versus WT Tissue
Apoptotic indexes were quantiﬁed on 6 retinas as described in Colombani
et al. (2006).
Standard Growth Conditions and Size Measurements
The experiment was performed as described in Genevet et al. (2009). Ten wing
discs for each genotype were analyzed.
Fly Eye Scanning Electron Microscopy
Scanning microscopy of adult ﬂies was performed as described in Polesello
et al. (2006).
RNAi Treatment and Coimmunoprecipitation of Proteins
For RNAi treatment, S2 or S2R+ cells were treated with dsRNAs as indicated
for 4 days. For coimmunoprecipitations, S2R+ or S2 cells transfected with the
indicated plasmids were used. See Supplemental Experimental Procedures
MS1096 > > (control) and MS1096 > > yki wing discs were dissected in PBS
and snap-frozen in liquid nitrogen. RNA isolation and subsequent qRT-PCR
reactions were performed as described in Genevet et al. (2009). See Supple-
mental Experimental Procedures for primer sequences.
All error bars displayed represent standard deviations. All statistical analyses
performed (except epistasis analysis) were assessed by Mann-Whitney
nonparametric tests using the website http://elegans.swmed.edu/leon/
The epistasis analysis was made by pairwise comparison after correction
for the batch effect on 42 to 310 ﬂies of each genotype, divided in 4 to 6
cohorts. The approach used was a three-way log-linear model, against a
null-hypothesis of no interaction between phenotype and population. The p
value indicates whether the pair of populations differ in their phenotype
proﬁles: p value(kibra
+ UAS ex) = 2.69 3 10
, p value(kibra
+UAS mer) = 6.07 3 10
, p value(ex
+ UAS kibra) = 1.76 3
Supplemental Information includes four ﬁgures, Supplemental Experimental
Procedures, and Supplemental References and can be found with this article
online at doi:10.1016/j.devcel.2009.12.011.
We thank D. Pan, R. Fehon, G. Halder, A. Laughon, K. Irvine, J. Kremer-
skothen, G. Struhl, T. Igaki, H. McNeill, the Bloomington and VDRC stock
centres, and the DGRC for ﬂy stocks and reagents. We are very grateful to
G. Dietzl, B. Dickson, and S. Cohen for their support in carrying out the RNAi
screen. We are grateful to K. Blight, A. Weston, and L. Collinson from the
LRI Electron Microscopy Facility and to P. Jordan from the Light Microscopy
Facility for technical help, to G. Kelly for help with statistics, to F. Josue
help with apoptotic index analysis, as well as to T. Gilbank, S. Murray, and
S. Maloney from the Fly Facility for technical support. We thank S. Cohen
and C. Polesello for discussions and comments on the manuscript. We thank
H. Stocker and E. Hafen for discussing data prior to publication. M.C.W. is sup-
ported by an EMBO long-term fellowship. The Tapon and Thompson laborato-
ries are supported by Cancer Research UK.
Received: May 27, 2009
Revised: October 20, 2009
Accepted: December 24, 2009
Published: February 15, 2010
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