Reagents

Stock #

Genotype

Allele type | insertion chromosome

Reference

wimp (RpII140 – RNA Polymerase II subunit)

5874

wimp, rucuca/TM3, Sb

antimorphic allele

1

Rho1 GTPase

9477

w*; b pr cn Rho1[1B] px sp/CyO

null allele

2,3,5

Cdc42  [R186C point mutation]

[Cdc42 point mutation (Cdc42(R186C)) under control of the UASp promoter (inducible)]
[This is a dominant negative Cdc42 mutation that gives similar phenotypes in flies to that observed in the equivalent human mutations.]

90925

w*  P{w+, UASp-Cdc42(R186C)}11

X

15

90926

w*; P{w+, UASp-Cdc42(R186C)}26/CyO

2nd

15

90927

w*; P{w+, UASp-Cdc42(R186C)}37

3rd

15

washout  (Wiskott-Aldrich Syndrome family protein)

79220

w*; wash[Delta185]

null allele,background lethal removed

14

28285

w*; wash[Delta185]/CyO

null allele

6,9

Sqh-mScarlet

[This is a CRISPR knock-in of mScarlet-i into the Spaghetti squash locus (C-terminal fusion)]

94929

TI{TI}sqh^mScarlet-I ; MKRS/TM6B

CRISPR knock-in (X chr)

19

sqh gRNA

[gRNA for sqh knock-in into pCFD5 vector]

94930

w*;  P{U6:3-sqh.gRNA.pCFD5}attp2

gRNA insertion on 3rd

19

degringolade

35525

w*; dgrn[DK]/TM3, Sb

null allele

9

"Red" balancers (sChFP : sqh driven, ChFP protein)

35522

FM7a, P{w+; sChFP}

X

11

35523

w*; Sco/CyO, P{w+, sChFP}

2nd

11

35524

w*; Gl/TM3, Sb P{w+, sChFP}    

3rd

11

photoconvertible actin binding domain

[sM3MCA : sqh driven, Maple3 fluorescent protein, moesin α-helical-coiled and actin binding site]  

94924

w*;  P{w+, sM3MCA}12 

X

19

94925

w*;  P{w+, sM3MCA}20/CyO; MKRS/TM3,Ser

2nd

19

94926

w*;  Kr/CyO; P{w+, sSM3MCA}33 

3rd

19

actin binding domain (Scarlet)

[sStMCA : sqh driven, Scarlet fluorescent protein, moesin α-helical-coiled and actin binding site]
[This Scarlet version gives much brighter fluorescence and less background than mCherry or Kate2. This reporter also recognizes nuclear actin.]

90928

w*  P{w+, sStMCA}10

X

16

90929

w*;  P{w+, sStMCA}20

2nd

16

90930

w*;  P{w+, sStMCA}30

3rd

16

actin binding domain (Cherry)

(sChMCA : sqh driven, ChFP protein, moesin α-helical-coiled and actin binding site)

35519

 w* P{w+, sChMCA}#12

X

10

35520

w*; P{w+, sChMCA}#22

2nd

10

35521

w*; P{w+, sChMCA}#31

3rd

10

actin binding domain (Kate2)

(sK2MCA: sqh driven, Kate2 fluorescent protein, moesin α-helical-coiled and actin binding site)
(Kate2 is supposed to be far red.  It isn’t in flies, but it is better separated from GFP than CherryFP (giving less background fluorescence)).

76268

w* P{w+, SK2MCA}#12

X

14

76269

w*;  P{w+, sK2MCA}#25

2nd

14

76270

w*;  P{w+, sK2MCA}#36

3rd

14

actin binding protein (Cyan)

[sTMCA: sqh driven, mTFP1 (cyan) fluorescent protein, moesin a-helical-coiled and actin binding site]

606720

w*  P{w+, sTMCA}10       

X chromo insertion

606721

w*; P{w+, sTMCA}30       

3^rd chromo insertion

actin (inducible RFP fusion protein)

24777

P{UASp-RFP.actin}13, w*

X

7

24778

w*; P{UASp-RFP.actin}29

2nd

7

24779

w*; P{UASp-RFP.actin}38

3rd

7

GCaMP6s

[This is a constitutive version of the GCaMP6s calcium reporter that works very well in embryos.]

91362

w[*]; P{w[+mC]=sqh-GCaMP6s]101.1 sqh-GCaMP6s]101.2

X    (two copies of transgene)

17

91363

w[*]; P{w[+mC]=sqh-GCaMP6s}20

2^nd  (one copy of transgene)

17

91364

w[*]; P{w[+mC]=sqh-GCaMP6s]305.1 sqh-GCaMP6s]305.2

3rd  (two copies of transgene)

17

GCaMP6s

[This is an inducible (UAS) version of the GCaMP6s calcium reporter in the pUASp vector (works well in the germline and embryo).]

91365

w[*]; P{w[+mC]=UASp-GCaMP6s}20/CyO

2nd

91366

w[*]; P{w[+mC]=UASp-GCaMP6s}30/TM3, Sb

3rd

Sep1 RNAi

(Express shRNA for Sep1 under UASz control)

604937

w* ; P{y[+t7.7]=UASz-Sep1-shRNA }JK22C

2nd

21

604938

w*;;  M{UASz-Sep1-shRNA }ZH-86Fb

3rd

21

AnxB10-mScarlet

(This is a CRIMIC knock-in of mScarlet-i (StFP) into the AnxB10 locus)

605570

y[1] w[*] TI{CRIMIC.StFP}AnxB10[CR00582-StFP]

X

22

sfGFP-AnxB10

(sfGFP-AnxB10 fusion protein under control of sqh promoter)

605571

w*, P{w[+mC]=sqh-sfGFP-AnxB10}10

X

22

605572

w*;  P{w[+mC]=sqh-sfGFP-AnxB10}20; MKRS/TM6B

2nd

22

605573

w*;;  P{w[+mC]=sqh-sfGFP-AnxB10}30

3rd

22

StFP-AnxB11

(mScarlet-i (StFP)-AnxB11 fusion protein under control of endogenous AnxB11 promoter)

605574

w*, P{StFP-AnxB11 }10

X

22

605575

w*; P{StFP-AnxB11 }20

2nd

22

605576

w*;; P{StFP-AnxB11 }31

3rd

22

AnxB11 RNAi

(Express shRNA for AnxB11 under Walium22 control)

605568

w*;;  M{UAS-AnxB11-shRNA1 }ZH-86Fb

3rd

22

605569

w*;;  M{UAS-AnxB11-shRNA2 }ZH-86Fb

3rd

22

StFP-8PM

(Plasma membrane specific reporter.  mScarlet-i (StFP) fused to prenylated octavalent that associates to plasma membrane under control of Ubiquitin promoter)

605577

w*, P{w[+mC]=Ubi-StFP-8PM }10

X

22

605578

w*;  P{w[+mC]= Ubi-StFP-8PM }25

2nd

22

605579

w*;;  P{w[+mC]= Ubi-StFP-8PM }31

3rd

22

Pav-sfGFP knock-in

[Knock-in allele of pav expressing Pav protein fused to super-folder GFP at C-terminus]

606719

w*, TI{TI}pav[sfGFP]                          

  3^rd chromo knock-in

sfGFP-Shrub

[super-folder GFP fused to N-terminus of Shrub expressed by sqh promoter]

606716

w*  P{w+, sqh-sfGFP-shrub}21                                     

2^nd chromo insertion

 606717

  w* sStMCA; ; P{w+, sqh-sfGFP-shrub }31       

3^rd chromo insertion

StFP-Shrub

[mScarlet-i fused to N-terminus of Shrub expressed by sqh promoter]

606718

  w*, sGMCA; ; P{w+, sqh-StFP-shrub }21        

2^nd chromo insertion

StFP-Lact-C2

(PS lipid biosensor.  mScarlet-i (StFP) fused to Lact-C2 domain that associates to phosphatidylserine under UASz control)

605580

w*;;  M{UASz-StFP-Lact-C2 }ZH-86Fb

3rd

22

PASS-sfGFP

(PA lipid biosensor.  sfGFP fused to PASS domain that associates to phosphatidic acid under UASz control)

605581

w*;;  M{UASz-PASS-sfGFP }ZH-86Fb

3rd

22

PASS-StFP

(PA lipid biosensor.  mScarlet-i (StFP) fused to PASS domain that associates to phosphatidic acid under UASz control)

605582

w*;;  M{UASz-PASS-StFP }ZH-86Fb

3rd

22

Arp2-sfGFP

(superfolder GFP fused C-terminal to full-length Arp2 and expressed under control of the Arp2 promoter).

605605

w*  P{w+, Arp2-sfGFP}11

X

605606

w*; P{w+, Arp2-sfGFP}20

2nd

Arp3-StFP

(mScarlet-i (StFP) fused C-terminal to full-length Arp3 and expressed under control of the Arp3 promoter).

605607

w*  P{w+, Arp3-StFP}10

X

605608

w*; P{w+, Arp3-StFP}20

2nd

605609

w*; P{w+, Arp3-StFP}30

3rd

Wash

[GFP fusion protein under control of the UASp promoter (inducible)]

81639

w*; P{w[+mC]=UASp-GFP-Wash}15

X

81640

w*; P{w[+mC]=UASp-GFP-Wash}26

2nd

81641

w*; P{w[+mC]=UASp-GFP-Wash}35

3rd

Wash

[CherryFP fusion protein under control of the UASp promoter (inducible)]

81642

w*; P{w[+mC]=UASp-ChFP-Wash}10

2nd

18

81643

w*; P{w[+mC]=UASp-ChFP-Wash}31

3rd

18

Wash

[super folder GFP fusion protein under the control of the endogenous Wash promoter]

81644

w*; P{w[+mC]=Wash-sfGFP-Wash}104

2nd

14

SCAR

[GFP-SCAR fusion protein under control of endogenous SCAR promoter]

94927

w*;  P{GFP- SCAR }20; Dr/TM3,Sb

2nd

19

94928

w*;  Sp/CyO; P{GFP- SCAR }30

3rd

19

Rho1 GTPase 

(GFP fusion protein under control of endogenous Rho1 promoter)

9527

P{Rho1.GFP}, w*

X

5

9528

w*; P{Rho1.GFP}       

2nd

5

24762

w*; P{Rho1.GFP}30

3rd

5

Rho1 GTPase 

(ChFP fusion protein under control of endogenous Rho1 promoter)

52280

P{w[+mC]=mChFP-Rho1}10, w[*]

X

13

52281

w[*]; P{w[+mC]=mChFP-Rho1}21     

2nd

13

52282

w[*]; sna[Sco]/CyO; P{w[+mC]=mChFP-Rho1}31

3rd

13

Cdc42 GTPase  (sfGFP)

[super-folder GFP protein fused to N-terminus of Cdc42 expressed by Cdc42 endogenous promoter]

606710

w*  P{w+, sfGFP-Cdc42}11       

X chromo insertion

606711

w*; P{w+, sfGFP-Cdc42}20       

2^nd chromo insertion

606712

w*; P{w+, sfGFP-Cdc42}33       

3^rd chromo insertion

Cdc42 GTPase (Cherry)

(ChFP fusion protein under the control of the Sqh promoter)

42236

w*;  P{w[+mC]=sqh-ChFP-Cdc42}23

2nd

11

42237

w*;  P{w[+mC]=sqh-ChFP-Cdc42}3

3rd

11

Rac1 GTPase 

(GFP fusion protein under control of endogenous Rac1 promoter)

52283  

P{w[+mC]=GFP-Rac1}10, w[*]

X

13

52284

w[*]; P{w[+mC]=GFP-Rac1}20

2nd

13

52285

w[*]; P{w[+mC]=GFP-Rac1}30

3rd

13

Rac1 GTPase 

(CherryFP fusion protein under control of endogenous Rac1 promoter)

76266

w[*];  P{w[+mC]=ChFP-Rac1}21

2nd

14

76267

w[*];  P{w[+mC]=ChFP-Rac1}30

3rd

14

Rac2 GTPase 

(GFP fusion protein under control of endogenous Rac2 promoter)

52286

w[*]; P{w[+mC]=GFP-Rac2}21

2nd

13

52287

w[*]; P{w[+mC]=GFP-Rac2}31

3rd

13

Tumbleweed

(super folder GFP fusion protein under the control of the Sqh promoter)

76264

w*;  P{w[+mC]=sqh-sfGFP-Tum}20

2nd

14

76265

w*;  P{w[+mC]=sqh-sfGFP-Tum}30

3rd

14

Pebble

(GFP fusion protein under the control of the Sqh promoter)

76257

w* P{w[+mC]=sqh-Pbl-eGFP}10

X

14

76258

w*;  P{w[+mC]=sqh-Pbl-eGFP}30

3rd

14

RhoGEF2 

(super folder GFP fusion protein under the control of the Sqh promoter)

76259

w* P{w[+mC]=sqh-sfGFP-RhoGEF2}10

X

14

76260

w*;  P{w[+mC]=sqh-sfGFP-RhoGEF2}30

3rd

14

RhoGEF3  

(CherryFP fusion protein under control of the UASp promoter (inducible))

76261

w*; P{w[+mC]=UASp-ChFP-RhoGEF3}30

3rd

14

RhoGEF3

(super folder GFP fusion protein under control of the UASp promoter (inducible))

76262

w*; P{w[+mC]=UASp-sfGFP-RhoGEF3}20

2nd

14

76263

w*; P{w[+mC]=UASp-sfGFP-RhoGEF3}30

3rd

14

RhoGEF2 RNAi

(21nt shRNA from RhoGEF2 cloned into WALLIUM22; 99% knockdown by qPCR)

76255

M{w[+mC]=UAS-RhoGEF2.shRNA}ZH-86Fb

3rd

14

RhoGEF3 RNAi

(21nt shRNA from RhoGEF3 cloned into WALLIUM22; 92% knockdown by qPCR)

76256

M{w[+mC]=UAS-RhoGEF3.shRNA}ZH-86Fb

3rd

14

Rok

(GFP fusion protein under control of Sqh promoter)

52288

P{w[+mC]=sqh-GFP-rok}10, w[*]

X

13

52289

w[*]; P{w[+mC]=sqh-GFP-rok}30

3rd

13

Pak1

(GFP fusion protein under control of the UASp promoter (inducible)

52299

w[*]; P{w[+mC]=UASp-GFP-Pak1}300

3rd

13

Sticky

[super folder GFP fusion protein under the control of the Sqh promoter (constitutive)]

81645

w*; P{w[+mC]=sqh-sfGFP-Sti}10

X

81646

w*; P{w[+mC]=sqh-sfGFP-Sti}20

2nd

81647

w*; P{w[+mC]=sqh-sfGFP-Sti}32

3rd

Sticky

[CherryFP fusion protein under control of the UASp promoter (inducible)]

81648

w*; P{w[+mC]=UASp-ChFP-Sti}20/CyO

2nd

81649

w*; P{w[+mC]=UASp-ChFP-Sti}30/TM3

3rd

Rho family GTPase activity biosensors

(reporters for active Rho1, Rac, and/or Cdc42).

Rok^RBD

GFP fusion to Rok’s Rho1/Rac binding domain (RBD) expressed under control of the UASp promoter (inducible).

52290

w[*]; P{w[+mC]=UASp-rok.RBD-GFP}30

3rd

13

Dia^RBD

GFP fusion to Diaphanous’s Rho1 binding domain (RBD) expressed under control of the UASp promoter (inducible).

52291

w[*]; P{w[+mC]=UASp-dia.RBD-GFP}20

2nd

13

52292

w[*]; P{w[+mC]=UASp-dia.RBD-GFP}37

3rd

13

Capu^RBD

GFP fusion to Cappuccino’s Rho1 binding domain (RBD) expressed under control of the UASp promoter (inducible).

52293

w[*]; P{w[+mC]=UASp-capu.RBD-GFP}26

2nd

13

52294

w[*]; P{w[+mC]=UASp-capu.RBD-GFP}34

3rd

13

Wash^RBD

GFP fusion to Wash’s Rho1 binding domain (RBD) expressed under control of the UASp promoter (inducible).

52295

w[*]; P{w[+mC]=UASp-wash.RBD-GFP}25

2nd

13

52296

w[*]; P{w[+mC]=UASp-wash.RBD-GFP}34

3rd

13

Pkn^RBD (Scarlet)

[Rho1/Rac GTPase activity biosensor. Constitutively expresses the Rho1/Rac binding domain of Pkn fused to mScarlet-i under control of sqh regulatory sequences]

606713

w*  P{w+, sqh-Pkn.RBD.G58A-StFP}10        

X chromo insertion

606714

w*; P{w+, sqh-Pkn.RBD.G58A-StFP }21       

2^nd chromo insertion

606715

w*; P{w+, sqh-Pkn.RBD.G58A-StFP }30

3^rd chromo insertion

Pkn^RBD (GFP)

GFP fusion to Pkn’s Rho/Rac binding domain (RBD) expressed under control of the Sqh promoter (constitutive).

52297

w[*]; P{w[+mC]=sqh-Pkn.RBD.G58A-eGFP}212a
and P{sqh-Pkn.RBD.G58A-eGFP}212b

2nd

13

52298

w[*]; P{w[+mC]=sqh-Pkn.RBD.G58A-eGFP}312a
and P{sqh-Pkn.RBD.G58A-eGFP}312b

3rd

13

WASp^RBD

GFP fusion to WASp’s Cdc42 binding domain (RBD) expressed under control of the Sqh promoter (constitutive).

56745

w[*]; P{w[+mC]=sqh-WASp.RBD-GFP}278a
P{sqh-WASp.RBD-GFP}278b/CyO

2nd

13

56746

w[*]; P{w[+mC]=sqh-WASp.RBD-GFP}378a
P{sqh-WASp.RBD-GFP}378b

3rd

13

Pak1^RBD

GFP fusion to Pak1’s Rac/Cdc42 binding domain (RBD) expressed under control of the Sqh promoter (constitutive) or under the control of the UASp promoter (conditional).

56549

w[*]; P{w[+mC]=sqh-Pak1.RBD-GFP}21

2nd

13

56550

w[*]; P{w[+mC]=sqh-Pak1.RBD-GFP}31/TM3, Sb[1]

3rd

13

56548

w[*]; P{w[+mC]=UASp-Pak.RBD-GFP}30/TM3, Sb[1]

3rd

13

Pak3^RBD

GFP fusion to Pak3’s Rac/Cdc42 binding domain (RBD) expressed under control of the Sqh promoter (constitutive).

52303

w[*]; P{w[+mC]=sqh-Pak3.RBD-GFP}20

2nd

13

52304

w[*]; P{w[+mC]=sqh-Pak3.RBD-GFP}30

3rd

13

Cappuccino

(inducible GFP fusion protein)

24763

P{UASp-GFP.Capu}10, w*

X

5

24764

w*; P{UASp-GFP.Capu}20

2nd

5

Spire – C isoform

(inducible GFP fusion protein)

24765

w*; P{UASp-GFP.SpireC}21

2nd

5

24766

w*; P{UASp-GFP.SpireC}32

3rd

5

Spire – D isoform

(inducible GFP fusion protein)

24767

w*; P{UASp-GFP.SpireD}30

3rd

5

sisyphus, unconventional myosin 10A (myosin XV homolog)

(inducible RFP or GFP fusion proteins)

24780

P{UASp-Myo10A.GFP}10, w*

X

7

24781

w*; P{UASp-Myo10A.GFP}20

2nd

7

24782

w*; P{UASp-Myo10A.GFP}30        

3rd

7

24783

P{UASp-Myo10A.mRFP}11, w* 

X

7

24784

w*; P{UASp-Myo10A.mRFP}22     

2nd

7

24785

w*; P{UASp-Myo10A.mRFP}33 

3rd

7