REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Rabbit anti-GFP	Gift from U. Mayor (U. Mayor, personal communication)	N/A
Chicken anti-GFP	Millipore	Cat# 06-896, RRID: AB_310288
Guinea pig anti-Dpn	Gift from J. Knoblich (Levy and Larsen, 2013)	RRID: AB_2314299
Guinea pig anti-Mob4	Gift from T. Littleton (Schulte et al., 2010)	N/A
Mouse anti-Dlg	DSHB	Cat# 4F3 anti-discs large, RRID: AB_528203
Rabbit anti-pH3	Abcam	Cat# ab5176, RRID: AB_304763
Rabbit anti-Cka	Gift from W. Du (Weng and Wei, 2002)	N/A
Rabbit anti-pAKTS505	CST	Cat# 4054, RRID: AB_331414
Mouse anti-Flag clone M2	Sigma	Cat# F3165, RRID: AB_259529
Rat anti-HA clone 3F10	Roche	Cat# 11867431001, RRID: AB_390919
Rat IgG	Sigma	Cat# I4131, RRID: AB_1163627
Rabbit anti-Akt	CST	Cat# 9272, RRID: AB_329827
Rabbit anti-b-Actin	CST	Cat# 4967, RRID: AB_330288
Mouse anti-Myc clone 9E10	Santa Cruz Biotechnology	Cat# sc-40, RRID: AB_627268
Rabbit anti-pMST1T183/pMST2T180	CST	Cat# 3681S, RRID: AB_330269
Guinea pig anti-Hippo	Gift from G. Halder (Hamaratoglu et al., 2006)	N/A
Chemicals, Peptides, and Recombinant Proteins
Okadaic Acid	CST	Cat# 5934
Critical Commercial Assays
Click-iT EdU Alexa Fluor 594 Imaging Kit	Invitrogen	Cat# C10339
Co-Immunoprecipitation Kit	Pierce	Cat# 26149
Deposited Data
Transcriptome data	This paper	GEO: GSE128646
Experimental Models: Cell Lines
D. melanogaster: Cell line S2R+	Gift from B. Houdsen	FlyBase: FBtc0000150
Experimental Models: Organisms/Strains
D. melanogaster: mob4 mutant: y[1] w[∗]; Mob4[EYDeltaL3]/CyO	Bloomington Drosophila Stock Center (Schulte et al., 2010)	BDSC: 36331; FlyBase: FBst0036331
D. melanogaster: UAS line expressing mob4 RNAi: P{UAS-Mob4.RNAi.JS1}attP2	Bloomington Drosophila Stock Center (Schulte et al., 2010)	BDSC:36488; FlyBase: FBst0036488
D. melanogaster: UAS line expressing mob4: y[1] v[1]; P{y[+t7.7] v[+t1.8] = UAS-Mob4.S}attP2	Bloomington Drosophila Stock Center (Schulte et al., 2010)	BDSC: 36329; FlyBase: FBst0036329
D. melanogaster: UAS line expressing hMOB4: y[1] v[1]; P{y[+t7.7] v[+t1.8] = UAS-phocein.1}attP2	Bloomington Drosophila Stock Center (Schulte et al., 2010)	BDSC: 36330; FlyBase: FBst0036330
D. melanogaster: UAS line expressing cka-eGFP: w[∗]; P{w[+mC] = UASp-Cka.EGFP.C}2	Bloomington Drosophila Stock Center	BDSC: 53756; FlyBase: FBst0053756
D. melanogaster: UAS line expressing wdb-DN: w[∗]; P{w[+mC] = UAS-wdb.95-524.HA}6	Bloomington Drosophila Stock Center (Hannus et al., 2002)	BDSC: 55053; FlyBase: FBst0055053
D. melanogaster: UAS line expressing wts RNAi: y[1] sc[∗] v[1]; P{y[+t7.7] v[+t1.8] = TRiP.GL01331}attP2	Bloomington Drosophila Stock Center (Ding et al., 2016)	BDSC: 41899; FlyBase: FBst0041899
D. melanogaster: UAS line expressing hpo RNAi: y[1] v[1]; P{y[+t7.7] v[+t1.8] = TRiP.HMS00006}attP2	Bloomington Drosophila Stock Center (Ding et al., 2016)	BDSC: 33614; FlyBase: FBst0033614
D. melanogaster: UAS line expressing cka RNAi: y[1] v[1]; P{y[+t7.7] v[+t1.8] = TRiP.HM05138}attP2	Bloomington Drosophila Stock Center	BDSC: 28927; FlyBase: FBst0028927
D. melanogaster: Wild type: Oregon-R	Gift from M. Akam (Bossing and Technau, 1994)	FlyBase: FBsn0000276
D. melanogaster: UAS line expressing rheb: w[∗]; P{w[+mC] = UAS-Rheb.Pa}3	Gift from R. Sousa-Nunes (Sousa-Nunes et al., 2011)	BDSC: 9689; FlyBase: FBst0009689
D. melanogaster: Gal4 line under the control of grh: Grh-Gal4	Gift from A.H. Brand (Chell and Brand, 2010)	N/A
D. melanogaster: UAS line expressing mts-DN-HA: UAS-mts.dn181-HA	Gift from S. Eaton (Hannus et al., 2002)	N/A
D. melanogaster: line expressing pleckstrin homology domain-GFP fusion protein: PH-GFP (tGPH)	Gift from B. Edgar (Britton and Edgar, 1998)	N/A
D. melanogaster: bantam GFP-sensor line, ban-sensor (db20)	Gift from S.M. Cohen (Brennecke et al., 2003)	N/A
D. melanogaster: mtsXE225839 mutant: mtsXE2258/CyO, P{sevRas1.V12}F1 and mts299 mutant	Gifts from H. Wang (Wang et al., 2009)	BDSC: 5684; FlyBase:FBst0005684 N/A for mts299
D. melanogaster: Gal4 line under the control of insc: w[∗]; P{w[+mW.hs] = GawB}insc[Mz1407]	Bloomington Drosophila Stock Center	BDSC: 8751; FlyBase:FBst0008751
D. melanogaster: Gal4 line under the control of repo: w[1118]; P{w[+m∗] = GAL4}repo/TM6, tb	Gift from A. Hidalgo	N/A
D. melanogaster: UAS line expressing CD8-GFP: y[1] w[∗]; P{w[+mC] = UAS-mCD8::GFP.L}LL5, P{UAS-mCD8::GFP.L}2	Lee and Luo, 1999	BDSC: 5137; FlyBase:FBst0005137
D. melanogaster: UAS line expressing dicer2: UAS-dicer2	Ding et al., 2016	N/A
Oligonucleotides
Primer: Anchored polyT AAGCAGTGGTATCAAC GCAGAGTACT(26)VN	Bossing et al., 2012	N/A
Primer: SM AAGCAGTGGTATCAACGCAGAG TACGCrGrGrG	Bossing et al., 2012	N/A
Primer: Nested AAGCAGTGGTATCAACGCAGAGT	Bossing et al., 2012	N/A
Primers used for RT-qPCR	See Table S4	N/A
dsRNA targeting sequence mob4: Forward: TAATAC GACTCACTATAGGGagatgtggaagtacgagcacctg	Schulte et al., 2010	N/A
dsRNA targeting sequence mob4: Reverse: TAATACG ACTCACTATAGGGagatgcgagaagatgcgatacac	Schulte et al., 2010	N/A
dsRNA targeting sequence cka: Forward: TAATACG ACTCACTATAGGGatacgggtccagttctgtgc	This paper	N/A
dsRNA targeting sequence cka: Reverse: TAATACG ACTCACTATAGGGtgttgtaggccaccacgata	This paper	N/A
dsRNA targeting sequence DsRed: Forward: TAATAC GACTCACTATAGGGgccgatgaacttcaccttgt	This paper	N/A
dsRNA targeting sequence DsRed: Reverse: TAATAC GACTCACTATAGGGcgaggacgtcatcaaggagt	This paper	N/A
Recombinant DNA
Plasmid: 12XCSL DsRedExpressDL	Hansson et al., 2006	Addgene plasmid #47683
Plasmid: Flag-NTAN	Gift from P. Ribeiro (Ribeiro et al., 2010)	N/A
Plasmid: Flag-Hippo	Gift from P. Ribeiro (Ribeiro et al., 2010)	N/A
Plasmid: Myc-Mts	Gift from P. Ribeiro (Ribeiro et al., 2010)	N/A
Plasmid: HA-Mts	Gift from P. Ribeiro (Ribeiro et al., 2010)	N/A
Plasmid: Myc-Akt	Gift from W. Hongyan (Li et al., 2014)	N/A
Software and Algorithms
R/Bioconductor Limma	Ritchie et al., 2015	https://bioconductor.org/packages/release/bioc/html/limma.html
STRING v10.5	Szklarczyk et al., 2015	https://string-db.org/cgi/input.pl?sessionId=QMsQ2cmXKFYZ&input_page_show_search=on
FlyAtlas	Chintapalli et al., 2007	http://flyatlas.org/atlas.cgi
DIOPT – DRSC Integrative Ortholog Prediction Tool	Hu et al., 2011	https://www.flyrnai.org/cgi-bin/DRSC_orthologs.pl