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. 2024 Jul 11;25(8):3324–3347. doi: 10.1038/s44319-024-00181-y

Reagents and tools table

Reagent/resource Reference or source Identifier or catalog number
Bacterial strains
E. coli DH5α Invitrogen 18265017
E. coli BL21(DE3) Merck Australia CMC0016
Antibodies
BNIP3 (ANa-40) mouse monoclonal Santa Cruz Biotechnology sc-56167
BNIP3 rabbit monoclonal Abcam ab109362
Cullin 1 rabbit Invitrogen 718700
FLAG monoclonal Sigma-Aldrich F3165
FLAG rabbit polyclonal Sigma-Aldrich SAB4301135
GFP Invitrogen MA5-15256
HA rabbit monoclonal Cell Signalling Technology 3724S
HDAC6 Santa Cruz Biotechnology sc-11420
HIF1α rabbit monoclonal Cell Signalling Technology 36169S
MTCO2 mouse monoclonal Abcam ab110258
MYC Bethyl Technologies A190-105A
NIX mouse monoclonal Santa Cruz Technologies sc-166332
NIX rabbit polyclonal Cell Signalling Technology 12396
p27 mouse monoclonal BD Biosciences 610242
PPTC7 rabbit Novus Biologicals NBP1-90654
Skp1 Pagano Laboratory
TOMM20 mouse monoclonal BD Biosciences Clone 29 612278
HSP60
Vinculin (VCL) mouse monoclonal Santa Cruz Biotechnology sc-55465
Donkey anti-mouse IgG Alexa FluorTM 488 Invitrogen A21202
Donkey anti-mouse IgG Alexa FluorTM 555 Invitrogen A31570
Donkey anti-mouse IgG Alexa Fluor TM 647 Invitrogen A31571
Donkey anti-rabbit IgG Alexa FluorTM 488 Invitrogen A21026
Donkey anti-rabbit IgG Alexa FluorTM 555 Invitrogen A31572
Donkey anti-rabbit IgG Alexa FluorTM 647 Invitrogen A31573
Chemicals
Benzamidine hydrochloride hydrate Sigma-Aldrich B6506
Deoxyribonuclease I (DNase I) Sigma-Aldrich DN25
Talon® resin Clontech 635503
Glutathione Sepharose 4B GE Healthcare GEHE17-0756-0
Isopropyl β-D-1-thiogalactopyranoside Bioline BIO-37036
Recombinant DNA
PPTC7-1_pLentiCRISPR v2 This paper
PPTC7-2_pLentiCRISPR v2 This paper
Fbxl4 CRISPR guide RNA 2_pSpCas9 BB-2A-Puro (PX459) v2.0 Nguyen-Dien
hPPTC7_pcDNA3.1(+)-C-DYK This paper
hPPTC7-pcDNA3.1-FLAG Natalie Niemi
pLV[Exp]-Puro-EF1A>hPPTC7[NM_139283.2]/HA This paper
FBXL4-pcDNA3.1-3XFLAG This paper
D290N_hPPTC7_pcDNA3.1(+)-C-DYK This paper
D223N_hPPTC7_pcDNA3.1(+)-C-DYK This paper
D78N_hPPTC7_pcDNA3.1(+)-C-DYK This paper
pLV_Puro_EF1A_PPTC7_FLAG This paper
D78N_pLV_Puro_EF1A_PPTC7_FLAG This paper
D223N_pLV_Puro_EF1A_PPTC7_FLAG This paper
D290N_pLV_Puro_EF1A_PPTC7_FLAG This paper
D78A_hPPTC7_pcDNA3.1(+)-C-DYK This paper
D78A/D223A_hPPTC7_pcDNA3.1(+)-C-DYK This paper
D223A_D78A_pLV_Puro_EF1A_PPTC7_HA This paper
D78A_pLV_Puro_EF1A_PPTC7_HA This paper
K130E_hPPTC7_pcDNA3.1(+)-C-DYK This paper
Q128R_hPPTC7_pcDNA3.1(+)-C-DYK This paper
Y179D_hPPTC7_pcDNA3.1(+)-C-DYK This paper
N181E_hPPTC7_pcDNA3.1(+)-C-DYK This paper
Q128R_pLV_Puro_EF1A_PPTC7_HA This paper
K130E_pLV_Puro_EF1A_PPTC7_HA This paper
Y179D_pLV_Puro_EF1A_PPTC7_HA This paper
N181E_pLV_Puro_EF1A_PPTC7_HA This paper
NIX-RPE-AAA This paper
NIXD140-150 This paper
M71E_pLV_FBXL4_HA_WT_VB3 This paper
Arg544E_pLV_FBXL4_HA_WT_VB3 This paper
M71E and Arg544E_pLV_FBXL4_HA_WT_VB3 This paper
pLVX TetOne BNIP3 This paper
pLVX TetOne NIX This paper
Plasmid: pGEX-6P-2 Cytiva 28-9545-50
Plasmid: pGEX-6P-2 GST-PPTC7(31-304) This study N/A
Plasmid: pGEX-6P-2 GST-PPTC7(31-304)(D290N) This study N/A
Deposited Data
SCFβTRPC (crystal structure) RSCB Protein Data Bank PDB ID: 6TTU
Human PPTC7 (protein sequence) Uniprot Q8NI37
Human FBXL4 (open reading frame) Uniprot Q9UKA2
Human BNIP3 (protein sequence) Uniprot Q12983
Human NIX (protein sequence Uniprot O60238
Software
Pymol Schrodinger, USA https://pymol.org/2/
Consurf Simpson et al, 2021 https://consurf.tau.ac.il/consurf_index.php
AlphaFold2 Multimer Sowter et al, 2001; Stojanovski et al, 2007 https://github.com/deepmind/alphafold
ColabFold and ColabFold batch Sun et al, 2024 https://github.com/sokrypton/ColabFold
Other
HiLoadTM Superdex75 PG GE Healthcare Catalogue: 28989333