Antibodies |
|
Mouse monoclonal anti-His |
QIAGEN |
Cat# 34660 |
Rabbit polyclonal anti-DAF-2 |
This Study |
N/A |
Rabbit polyclonal anti-CHN-1 |
This Study |
N/A |
Mouse monoclonal anti-FLAG |
Sigma-Aldrich |
Cat# F3165; RRID: AB_259529
|
Rabbit anti-UNC-45 |
Gazda et al., 2013 |
N/A |
Mouse Monoclonal anti-alpha Tubulin (Clone DM 1A) |
Sigma-Aldrich |
Cat# T9026; RRID: AB_477593
|
Mouse Monoclonal anti-alpha Tubulin (Clone B-5-1-2) |
Sigma-Aldrich |
Cat# T6074; RRID: AB_477582
|
Peroxidase-conjugated AffiniPure Goat Anti-Mouse IgG+IgM |
Jackson Immuno Research |
Cat# 115-035-003; RRID: AB_10015289
|
Peroxidase-conjugated AffiniPure Mouse Anti-Rabbit IgG+IgM |
Jackson Immuno Research |
Cat# 211-035-109; RRID: AB_2339150
|
Donkey anti-rabbit IRDye 800CW/680 |
Li-Cor |
Cat# 926-32223; RRID: AB_621845
|
Donkey anti-mouse IRDye 800CW/680 |
Li-Cor |
Cat# 926-32212; RRID: AB_621847
|
Mouse monoclonal anti-Actin |
Abcam |
Cat# ab8224; RRID: AB_449644
|
Rabbit polyclonal anti-AKT |
Cell Signaling |
Cat# 9272; RRID:AB_329827
|
Rabbit monoclonal anti-Phospho AKT (Ser473) |
Cell Signaling |
Cat# 4060S; RRID: AB_2315049
|
Rabbit polyclonal anti-phospho Drosophila AKT (Ser505) |
Cell Signaling |
Cat# 4054S; RRID: AB_331414
|
Rabbit monoclonal anti-ATG7 |
Novus Biologicals |
Cat# NBP1-95872; RRID: AB_11011190
|
Rabbit polyclonal anti-Chicken IgG |
Sigma-Aldrich |
Cat# C2288; RRID: AB_258791
|
Rabbit polyclonal anit-CHIP |
Milipore |
Cat# PC711; RRID: AB_2198058
|
Rat monoclonal anti-GFP |
Chromotek |
Cat# 3h9, ABIN398304; RRID: AB_10773374
|
Mouse monoclonal anti-Histidine-Tag |
Serotec |
Cat# MCA1396; RRID: AB_322084
|
Rat monoclonal anti-Hsp90 |
Abcam |
Cat# ab13494; RRID: AB_300398
|
Rabbit polyclonal anti-LC3B |
Novus Biologicals |
Cat# NB100-2220; RRID: AB_10003146
|
Mouse monoclonal anti-p62 |
Santa Cruz |
Cat# sc-28359; RRID: AB_628279
|
Rabbit anti-PDK1 |
Cell Signaling |
Cat# 3062; RRID: AB_2236832
|
Rabbit polyclonal anti-mTOR |
Abcam |
Cat# ab45989; RRID: AB_944296
|
Rabbit monoclonal anti-IGF1R |
Cell Signaling |
Cat# 9750S; RRID: AB_10950969
|
Rabbit polyclonal anti-INSR |
Santa Cruz Biotechnology |
Cat# sc-711; RRID: AB_631835
|
Goat polyclonal anti-gamma Tubulin |
Santa Cruz Biotechnology |
Cat# sc-7396; RRID: AB_2211262
|
Rabbit polyclonal anti-Stub1 (CHIP) |
Abcam |
Cat# 69451; RRID: AB_2198071
|
Rabbit polyclonal anti-dINSR |
This Study |
N/A |
|
Bacterial and Virus Strains |
|
BL21-CodonPlus (DE3)-RIL |
New England Biolabs |
Cat# C2530H |
OP50 |
CGC |
N/A |
HT115 |
CGC |
N/A |
|
Chemicals, Peptides, and Recombinant Proteins |
|
completeTablets EDTA free |
Sigma-Aldrich |
Cat# 4693132001 |
UbcH5b |
Boston Biochem |
Cat# E2-622 |
FLAG::Ubiquitin |
Boston Biochem |
Cat# U-120 |
E1 activating Enzyme |
Enzo Life Sciences |
Cat# BML-UW9410-0050 |
Ubiquitin Conjugation Reaction Buffer |
Enzo Life Sciences |
Cat# BML-KW9885-0001 |
ATP Regeneration Solution |
Enzo Life Sciences |
Cat# BML-EW9810-0100 |
Ubiquitin |
Sigma-Aldrich |
Cat# U6253 |
UbK0 |
Boston Biochem |
Cat# UM-Nok |
FLAG M2 Affinity Gel |
Sigma-Aldrich |
Cat# A2220 |
3X FLAG peptide |
Sigma-Aldrich |
Cat# F4799 |
Bafilomycin A1 |
LC-Labs |
Cat# B1080 |
Chloroquine diphosphate salt |
Sigma-Aldrich |
Cat# C6628 |
Dynamin Inhibitor V-34-2 |
Calbiochem |
Cat# 324414 |
Methylviologen dichloride hydrate |
Sigma-Aldrich |
Cat# 856177-CAS75365-73-0 |
MG132 |
Peptanova |
Cat# 3175-V |
L-Canavanine |
Sigma-Aldrich |
Cat# C1625 |
Paraquat dichloride hydrate |
Sigma-Aldrich |
Cat# 36541 |
RU486 (Mifepristone) |
Sigma-Aldrich |
Cat# M8046 |
VSV::CHN-1::6His |
This Study |
N/A |
VSV::CHN-1 H218Q::6His |
This Study |
N/A |
VSV::CHN-1 ΔU-box::6His |
This Study |
N/A |
VSV: CHN-1 N9D, K13A, K73A::6His |
This Study |
N/A |
UFD-2::6His |
This Study |
N/A |
6His::DAF-2 C terminus (aa 1205-1928) |
This Study |
N/A |
WWP-1::6His |
This Study |
N/A |
CHIP |
Arndt et al., 2005 |
N/A |
INSR Full Length |
This Study |
N/A |
INSR K1047R |
This Study |
N/A |
dINSR (aa 1815-2144) |
This Study |
N/A |
INSR C terminus (aa 987-1428) |
This Study |
N/A |
TRIzol |
Invitrogen |
Cat# 15596026 |
|
Critical Commercial Assays |
|
ECL kit |
GE Healthcare |
Cat# RPN2232 |
QuickChange Lightning Site-Directed Mutagenesis Kit |
Agilent Technologies |
Cat# 210519 |
InviTrap Spin Universal RNA Mini Kit |
Stratec |
Cat# 1060100300 |
jetPRIME transfection reagent |
Peqlab |
Cat# 114-75 |
High-Capacity cDNA Reverse Transcription Kit |
Applied Biosystems |
Cat# 4368814 |
Brilliant III Ultra-Fast SYBR Green QPCR Master Mix |
Agilent Technologies |
Cat# 600882 |
Sso Fast EvaGreen supermix |
Bio-Rad |
Cat# 1725201 |
iScript cDNA Synthesis Kit |
Bio-Rad |
Cat# 1708891 |
RNeasy Mini Kit |
QIAGEN |
Cat# 74104 |
|
Experimental Models: Cell Lines |
|
HEK293 (Human embryonic kidney cells) |
Sigma-Aldrich |
Cat# 85120602 |
HeLa (Epitheloid cervix carcinoma cells (female)) |
Sigma-Aldrich |
Cat# 93021013 |
|
Experimental Models: Organisms/Strains |
|
C. elegans: unc-119(ed4)III; hhIs136[unc-119(+); chn-1p::chn-1::FLAG]
|
This study |
N/A |
C. elegans:unc-119(ed4)III; hhIs145[unc-119(+); chn-1p::chn-1(U-box)::FLAG]
|
This study |
N/A |
C. elegans: Bristol (N2) strain as wild-type (WT) |
CGC |
N2 |
C. elegans: chn-1(by155)I
|
CGC |
WormBase ID: WBVar00000641 |
C. elegans:chn-1(tm2692)I
|
CGC |
WormBase ID: WBVar00251538 |
C. elegans:daf-16(mgDf47)I
|
CGC |
WormBase ID: WBVar00275574 |
C. elegans:daf-16(m26)I
|
CGC |
WormBase ID: WBVar00088538 |
C. elegans:ufd-2(tm1380)II
|
CGC |
WormBase ID: WBVar00250374 |
C. elegans:daf-2(e1370)III
|
CGC |
WormBase ID: WBVar00143949 |
C. elegans:daf-2(e1368)III
|
CGC |
WormBase ID: WBVar00143947 |
C. elegans:daf-2(m212)III
|
CGC |
WormBase ID: DR608 |
C. elegans:daf-2(m596)III
|
CGC |
WormBase ID: WBVar00088714 |
C. elegans:daf-2(m579)III
|
CGC |
WormBase ID: WBVar00088710 |
C. elegans:daf-2(sa193)III
|
CGC |
WormBase ID: WBVar00242513 |
C. elegans:daf-2(e979)III
|
CGC |
WormBase ID: WBVar00143653 |
C. elegans:daf-2(e1391)III
|
CGC |
WormBase ID: WBVar00143967 |
C. elegans:daf-2(m577)III
|
CGC |
WormBase ID: WBVar00088709 |
C. elegans:daf-2(gk390525)III
|
CGC |
WormBase ID:WBVar00813221 |
C. elegans:eat-2(ad465)
|
CGC |
WormBase ID: WBVar00000014 |
C.elegans: unc-119(ed4)III
|
CGC |
WormBase ID: WBVar00145094 |
C. elegans:dgEx80[pAMS66 vha-6p::Q44::YFP + rol-6(su1006)]
|
Mohri-Shiomi and Garsin, 2008 |
N/A |
C. elegans:zIs356(daf-16p::daf-16::GFP; rol-6)
|
Lee et al., 2001 |
N/A |
C. elegans:rmIs133(unc-54::Q40::YFP)
|
Morley et al., 2002 |
N/A |
D. melanogaster: w1118
|
Bloomington Stock Center |
RRID: BDSC_3605 |
D. melanogaster: actin-Gal4 |
Bloomington Stock Center |
RRID: BDSC_3954 |
D. melanogaster: RNAi-dCHIP |
VDRC |
RRID: FlyBase_FBst0460471 |
D. melanogaster: TIGS-2 |
Scott Pletscher |
N/A |
D. melanogaster: UAS-dCHIP |
Verena Arndt |
N/A |
|
Oligonucleotides |
|
See Table S3 for primer sequences |
This Study |
N/A |
siRNA human CHIP 5 |
QIAGEN |
Cat# SI00081963 |
siRNA human CHIP 5 |
QIAGEN |
Cat# SI03109659 |
siRNA human ATG7 1 |
QIAGEN |
Cat# SI04194883 |
siRNA human ATG7 3 |
QIAGEN |
Cat# SI04231360 |
Allstars negative control siRNA |
QIAGEN |
Cat# SI03650318 |
|
Recombinant DNA |
|
pET21a-VSV::CHN-1::6His |
This Study |
N/A |
pET21a- VSV::CHN-1 H218Q::6His |
This Study |
N/A |
pET21a- VSV::CHN-1 ΔU-box::6His |
This Study |
N/A |
pET21a- VSV: CHN-1 N9D, K13A, K73A::6His |
This Study |
N/A |
pET21a-UFD-2::6His |
This Study |
N/A |
pT7-6His::DAF-2 C terminus (aa 1205-1928) |
This Study |
N/A |
pET21a-WWP-1::6His |
This Study |
N/A |
pCMV-tag2b-CHIP |
Arndt et al., 2005 |
N/A |
pCMV-tag2b-INSR Full Length |
This Study |
N/A |
pCMV-tag2b-INSR K1047R |
This Study |
N/A |
pETM-11-dINSR (aa 1815-2144) |
This Study |
N/A |
pMAL-C2-INSR C terminus (aa 987-1428) |
This Study |
N/A |
pEGFP-polyQ-25 |
Westhoff et al., 2005 |
N/A |
pEGFP-polyQ-103 |
Westhoff et al., 2005 |
N/A |
|
Software and Algorithms |
|
GraphPad Prism 5 |
GraphPad Software |
https://www.graphpad.com/scientific-software/prism/ |
Image Studio 4.0 |
LI-COR Biosciences |
https://www.licor.com/bio/products/software/image_studio/ |
Excel 2013 |
Microsoft |
https://products.office.com/en-us/excel |
XLSTAT |
XLSTAT |
https://www.xlstat.com/en/ |
ImageJ 1.48v |
Wayne Rasband (NIH) |
https://imagej.nih.gov/ij/ |