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prk7
Prk7, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/prk7+flag/pRK7-FLAG-TSC2+(Plasmid+%238996)/pmc12839532-35-1-9
Average 93 stars, based on 12 article reviews
Prk7, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/prk7+flag/pRK7-FLAG-TSC2+(Plasmid+%238996)/pmc12839532-35-1-9
Average 93 stars, based on 12 article reviews
prk7 - by Bioz Stars,
2026-11
93/100 stars
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shRNA:Article Title: HUWE1 stimulates mTORC1 activity by enhancing Rheb interaction with mTORC1 and supports de novo pyrimidine synthesis. Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Mouse monoclonal anti-HA (clone 16B12) Covance Cat#MMS-101R; RRID:AB_291262 Mouse monoclonal anti-ubiquitinylated proteins (FK2) MilliporeSigma Cat#04–263; RRID:AB_612093 Alexa Fluor 594-goat polyclonal anti-rabbit IgG Thermo Fisher Scientific Cat#A11012; RRID:AB_141359 Alexa Fluor 488-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#A11029; RRID:AB_138404 Alexa Fluor 594-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#R37121; RRID:AB_2556549 Anti-rabbit IgG HRP-Linked Whole Ab GE Healthcare Cat#NA934; RRID:AB_772206 Anti-mouse IgG HRP-Linked Whole Ab GE Healthcare Cat#NA931; RRID:AB_772210 Bacterial and virus strains One Shot TM Stbl TM Chemically Competent E. coli Thermo Fisher Scientific Cat#C737303 Chemicals, peptides, and recombinant proteins Amino acid mixtures Sigma-Aldrich Cat#R7131 L-Glutathione reduced Sigma-Aldrich Cat#G4251 N-Ethylmaleimide (NEM) Sigma-Aldrich Cat#E3876 Saponin Sigma-Aldrich Cat#S4521 Actinomycin D Sigma-Aldrich Cat#A9415 Lipofectamine 3000 Invitrogen Cat#L3000001 Amino acid-free DMEM medium US Biological Life Sciences Cat#D9800-27 Leucine-free DMEM medium Crystalgen Cat#226-024 L-leucine RPI Cat#L22000 Dialyzed Fetal Bovine Serum Thermo Fisher Scientific Cat#26400044 Pierce TM Glutathione Agarose Thermo Fisher Scientific Cat#16100 Prolong Gold antifade reagent with DAPI Thermo Fisher Scientific Cat#P36931 Protein G Sepharose 4 Fast Flow GE Healthcare Cat#17061801 Flag peptide LifeTein Cat#LT12022 Insulin Sigma-Aldrich Cat#10908 Phosphorus-32 NEN Radiochemicals Cat#NEX011 MgATP Solution R&D Systems Cat#B-20 Critical commercial assays Subcellular Protein Fractionation Kit Thermo Fisher Scientific Cat#78840 TriFECTa RNAi Kits Integrated DNA Technologies (IDT) N/A Experimental models: Cell lines Human: HEK293T ATCC Cat#CRL-3216 Human: HeLa Yoshida et al. 71 N/A Human: HepG2 Sakasai-Sakai et al. 72 N/A Mouse: embryonic fibroblasts Yoshida et al. 71 N/A Mouse: embryonic fibroblasts p53 − /− Dr. David J Kwiatkowski 46 N/A Experimental models: Organisms/strains B6.129P2(Cg)-Huwe1 tm1.1Mak /J The Jaxon Laboratory Hao et al. 73 Strain#029254; RRID:IMSR_JAX:029254 B6.Cg-Speer6-ps1 Tg(Alb-cre)21Mgn /J The Jaxon Laboratory Postic et al. 74 Strain#003574; RRID:IMSR_JAX:003574 Oligonucleotides shRNA targeting HUWE1-1 (H and M) 5 ′ -GCATCTGTACAGTTCCATAGA This paper N/A shRNA targeting HUWE1-2 (H and M) 5 ′ -TGCCGCAATCCAGACATAT This paper N/A (Continued on next page) 18 Cell Reports 44, 116382, October 28, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER shRNA targeting HUWE1-3 (H and M) 5 ′ -GCTCAATACTAGCCGTCTACA This paper N/A shRNA targeting HUWE1-4 (H) 5 ′ -GGGAATTTCCCACCAGGATGA This paper N/A sgRNA targeting HUWE1 (H) 5 ′ -GCTGAACTTCACCTGAATCG This paper N/A sgRNA targeting GFP 5 ′ -CTTGTCACTACTTTCGGTTA Yao et al. 20 N/A shRNA targeting CAD-1 (M) 5 ′ -GCTCAGATCCTAGTGCTAACG This paper N/A shRNA targeting CAD-2 (M) 5 ′ -GGTGGTGATGGACATCTATGA This paper N/A siRNA sequences for HUWE1, see Table S1 Integrated DNA Technologies (IDT) N/A RT-qPCR primer sequences, see Table S2 This paper N/A Recombinant DNA pLKO1-Sc (Scramble shRNA) Sarbassov et al. 75 Addgene plasmid #1864 Lenti-CRISPR v2 Sanjana et al. 76 Addgene plasmid #52961 psPAX2 Dr. Didier Trono Addgene plasmid #12260 pMD2.G Dr. Didier Trono Addgene plasmid #12259 pcDNA3-HA-Ubiquitin Kamitani et al. 77 Addgene plasmid #18712 pcDNA3-Flag-HA-CAD Ben-Sahra et al. 31 Addgene plasmid #46238 pENTR1A-HUWE1 Dr. Jeanette G Cook Addgene plasmid #37431 pENTR1A-HUWE1 C4341S Dr. Jeanette G Cook Addgene plasmid # Quantitative RT-PCR:Article Title: HUWE1 stimulates mTORC1 activity by enhancing Rheb interaction with mTORC1 and supports de novo pyrimidine synthesis. Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Mouse monoclonal anti-HA (clone 16B12) Covance Cat#MMS-101R; RRID:AB_291262 Mouse monoclonal anti-ubiquitinylated proteins (FK2) MilliporeSigma Cat#04–263; RRID:AB_612093 Alexa Fluor 594-goat polyclonal anti-rabbit IgG Thermo Fisher Scientific Cat#A11012; RRID:AB_141359 Alexa Fluor 488-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#A11029; RRID:AB_138404 Alexa Fluor 594-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#R37121; RRID:AB_2556549 Anti-rabbit IgG HRP-Linked Whole Ab GE Healthcare Cat#NA934; RRID:AB_772206 Anti-mouse IgG HRP-Linked Whole Ab GE Healthcare Cat#NA931; RRID:AB_772210 Bacterial and virus strains One Shot TM Stbl TM Chemically Competent E. coli Thermo Fisher Scientific Cat#C737303 Chemicals, peptides, and recombinant proteins Amino acid mixtures Sigma-Aldrich Cat#R7131 L-Glutathione reduced Sigma-Aldrich Cat#G4251 N-Ethylmaleimide (NEM) Sigma-Aldrich Cat#E3876 Saponin Sigma-Aldrich Cat#S4521 Actinomycin D Sigma-Aldrich Cat#A9415 Lipofectamine 3000 Invitrogen Cat#L3000001 Amino acid-free DMEM medium US Biological Life Sciences Cat#D9800-27 Leucine-free DMEM medium Crystalgen Cat#226-024 L-leucine RPI Cat#L22000 Dialyzed Fetal Bovine Serum Thermo Fisher Scientific Cat#26400044 Pierce TM Glutathione Agarose Thermo Fisher Scientific Cat#16100 Prolong Gold antifade reagent with DAPI Thermo Fisher Scientific Cat#P36931 Protein G Sepharose 4 Fast Flow GE Healthcare Cat#17061801 Flag peptide LifeTein Cat#LT12022 Insulin Sigma-Aldrich Cat#10908 Phosphorus-32 NEN Radiochemicals Cat#NEX011 MgATP Solution R&D Systems Cat#B-20 Critical commercial assays Subcellular Protein Fractionation Kit Thermo Fisher Scientific Cat#78840 TriFECTa RNAi Kits Integrated DNA Technologies (IDT) N/A Experimental models: Cell lines Human: HEK293T ATCC Cat#CRL-3216 Human: HeLa Yoshida et al. 71 N/A Human: HepG2 Sakasai-Sakai et al. 72 N/A Mouse: embryonic fibroblasts Yoshida et al. 71 N/A Mouse: embryonic fibroblasts p53 − /− Dr. David J Kwiatkowski 46 N/A Experimental models: Organisms/strains B6.129P2(Cg)-Huwe1 tm1.1Mak /J The Jaxon Laboratory Hao et al. 73 Strain#029254; RRID:IMSR_JAX:029254 B6.Cg-Speer6-ps1 Tg(Alb-cre)21Mgn /J The Jaxon Laboratory Postic et al. 74 Strain#003574; RRID:IMSR_JAX:003574 Oligonucleotides shRNA targeting HUWE1-1 (H and M) 5 ′ -GCATCTGTACAGTTCCATAGA This paper N/A shRNA targeting HUWE1-2 (H and M) 5 ′ -TGCCGCAATCCAGACATAT This paper N/A (Continued on next page) 18 Cell Reports 44, 116382, October 28, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER shRNA targeting HUWE1-3 (H and M) 5 ′ -GCTCAATACTAGCCGTCTACA This paper N/A shRNA targeting HUWE1-4 (H) 5 ′ -GGGAATTTCCCACCAGGATGA This paper N/A sgRNA targeting HUWE1 (H) 5 ′ -GCTGAACTTCACCTGAATCG This paper N/A sgRNA targeting GFP 5 ′ -CTTGTCACTACTTTCGGTTA Yao et al. 20 N/A shRNA targeting CAD-1 (M) 5 ′ -GCTCAGATCCTAGTGCTAACG This paper N/A shRNA targeting CAD-2 (M) 5 ′ -GGTGGTGATGGACATCTATGA This paper N/A siRNA sequences for HUWE1, see Table S1 Integrated DNA Technologies (IDT) N/A RT-qPCR primer sequences, see Table S2 This paper N/A Recombinant DNA pLKO1-Sc (Scramble shRNA) Sarbassov et al. 75 Addgene plasmid #1864 Lenti-CRISPR v2 Sanjana et al. 76 Addgene plasmid #52961 psPAX2 Dr. Didier Trono Addgene plasmid #12260 pMD2.G Dr. Didier Trono Addgene plasmid #12259 pcDNA3-HA-Ubiquitin Kamitani et al. 77 Addgene plasmid #18712 pcDNA3-Flag-HA-CAD Ben-Sahra et al. 31 Addgene plasmid #46238 pENTR1A-HUWE1 Dr. Jeanette G Cook Addgene plasmid #37431 pENTR1A-HUWE1 C4341S Dr. Jeanette G Cook Addgene plasmid # Recombinant:Article Title: HUWE1 stimulates mTORC1 activity by enhancing Rheb interaction with mTORC1 and supports de novo pyrimidine synthesis. Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Mouse monoclonal anti-HA (clone 16B12) Covance Cat#MMS-101R; RRID:AB_291262 Mouse monoclonal anti-ubiquitinylated proteins (FK2) MilliporeSigma Cat#04–263; RRID:AB_612093 Alexa Fluor 594-goat polyclonal anti-rabbit IgG Thermo Fisher Scientific Cat#A11012; RRID:AB_141359 Alexa Fluor 488-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#A11029; RRID:AB_138404 Alexa Fluor 594-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#R37121; RRID:AB_2556549 Anti-rabbit IgG HRP-Linked Whole Ab GE Healthcare Cat#NA934; RRID:AB_772206 Anti-mouse IgG HRP-Linked Whole Ab GE Healthcare Cat#NA931; RRID:AB_772210 Bacterial and virus strains One Shot TM Stbl TM Chemically Competent E. coli Thermo Fisher Scientific Cat#C737303 Chemicals, peptides, and recombinant proteins Amino acid mixtures Sigma-Aldrich Cat#R7131 L-Glutathione reduced Sigma-Aldrich Cat#G4251 N-Ethylmaleimide (NEM) Sigma-Aldrich Cat#E3876 Saponin Sigma-Aldrich Cat#S4521 Actinomycin D Sigma-Aldrich Cat#A9415 Lipofectamine 3000 Invitrogen Cat#L3000001 Amino acid-free DMEM medium US Biological Life Sciences Cat#D9800-27 Leucine-free DMEM medium Crystalgen Cat#226-024 L-leucine RPI Cat#L22000 Dialyzed Fetal Bovine Serum Thermo Fisher Scientific Cat#26400044 Pierce TM Glutathione Agarose Thermo Fisher Scientific Cat#16100 Prolong Gold antifade reagent with DAPI Thermo Fisher Scientific Cat#P36931 Protein G Sepharose 4 Fast Flow GE Healthcare Cat#17061801 Flag peptide LifeTein Cat#LT12022 Insulin Sigma-Aldrich Cat#10908 Phosphorus-32 NEN Radiochemicals Cat#NEX011 MgATP Solution R&D Systems Cat#B-20 Critical commercial assays Subcellular Protein Fractionation Kit Thermo Fisher Scientific Cat#78840 TriFECTa RNAi Kits Integrated DNA Technologies (IDT) N/A Experimental models: Cell lines Human: HEK293T ATCC Cat#CRL-3216 Human: HeLa Yoshida et al. 71 N/A Human: HepG2 Sakasai-Sakai et al. 72 N/A Mouse: embryonic fibroblasts Yoshida et al. 71 N/A Mouse: embryonic fibroblasts p53 − /− Dr. David J Kwiatkowski 46 N/A Experimental models: Organisms/strains B6.129P2(Cg)-Huwe1 tm1.1Mak /J The Jaxon Laboratory Hao et al. 73 Strain#029254; RRID:IMSR_JAX:029254 B6.Cg-Speer6-ps1 Tg(Alb-cre)21Mgn /J The Jaxon Laboratory Postic et al. 74 Strain#003574; RRID:IMSR_JAX:003574 Oligonucleotides shRNA targeting HUWE1-1 (H and M) 5 ′ -GCATCTGTACAGTTCCATAGA This paper N/A shRNA targeting HUWE1-2 (H and M) 5 ′ -TGCCGCAATCCAGACATAT This paper N/A (Continued on next page) 18 Cell Reports 44, 116382, October 28, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER shRNA targeting HUWE1-3 (H and M) 5 ′ -GCTCAATACTAGCCGTCTACA This paper N/A shRNA targeting HUWE1-4 (H) 5 ′ -GGGAATTTCCCACCAGGATGA This paper N/A sgRNA targeting HUWE1 (H) 5 ′ -GCTGAACTTCACCTGAATCG This paper N/A sgRNA targeting GFP 5 ′ -CTTGTCACTACTTTCGGTTA Yao et al. 20 N/A shRNA targeting CAD-1 (M) 5 ′ -GCTCAGATCCTAGTGCTAACG This paper N/A shRNA targeting CAD-2 (M) 5 ′ -GGTGGTGATGGACATCTATGA This paper N/A siRNA sequences for HUWE1, see Table S1 Integrated DNA Technologies (IDT) N/A RT-qPCR primer sequences, see Table S2 This paper N/A Recombinant DNA pLKO1-Sc (Scramble shRNA) Sarbassov et al. 75 Addgene plasmid #1864 Lenti-CRISPR v2 Sanjana et al. 76 Addgene plasmid #52961 psPAX2 Dr. Didier Trono Addgene plasmid #12260 pMD2.G Dr. Didier Trono Addgene plasmid #12259 pcDNA3-HA-Ubiquitin Kamitani et al. 77 Addgene plasmid #18712 pcDNA3-Flag-HA-CAD Ben-Sahra et al. 31 Addgene plasmid #46238 pENTR1A-HUWE1 Dr. Jeanette G Cook Addgene plasmid #37431 pENTR1A-HUWE1 C4341S Dr. Jeanette G Cook Addgene plasmid # Plasmid Preparation:Article Title: HUWE1 stimulates mTORC1 activity by enhancing Rheb interaction with mTORC1 and supports de novo pyrimidine synthesis. Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Mouse monoclonal anti-HA (clone 16B12) Covance Cat#MMS-101R; RRID:AB_291262 Mouse monoclonal anti-ubiquitinylated proteins (FK2) MilliporeSigma Cat#04–263; RRID:AB_612093 Alexa Fluor 594-goat polyclonal anti-rabbit IgG Thermo Fisher Scientific Cat#A11012; RRID:AB_141359 Alexa Fluor 488-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#A11029; RRID:AB_138404 Alexa Fluor 594-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#R37121; RRID:AB_2556549 Anti-rabbit IgG HRP-Linked Whole Ab GE Healthcare Cat#NA934; RRID:AB_772206 Anti-mouse IgG HRP-Linked Whole Ab GE Healthcare Cat#NA931; RRID:AB_772210 Bacterial and virus strains One Shot TM Stbl TM Chemically Competent E. coli Thermo Fisher Scientific Cat#C737303 Chemicals, peptides, and recombinant proteins Amino acid mixtures Sigma-Aldrich Cat#R7131 L-Glutathione reduced Sigma-Aldrich Cat#G4251 N-Ethylmaleimide (NEM) Sigma-Aldrich Cat#E3876 Saponin Sigma-Aldrich Cat#S4521 Actinomycin D Sigma-Aldrich Cat#A9415 Lipofectamine 3000 Invitrogen Cat#L3000001 Amino acid-free DMEM medium US Biological Life Sciences Cat#D9800-27 Leucine-free DMEM medium Crystalgen Cat#226-024 L-leucine RPI Cat#L22000 Dialyzed Fetal Bovine Serum Thermo Fisher Scientific Cat#26400044 Pierce TM Glutathione Agarose Thermo Fisher Scientific Cat#16100 Prolong Gold antifade reagent with DAPI Thermo Fisher Scientific Cat#P36931 Protein G Sepharose 4 Fast Flow GE Healthcare Cat#17061801 Flag peptide LifeTein Cat#LT12022 Insulin Sigma-Aldrich Cat#10908 Phosphorus-32 NEN Radiochemicals Cat#NEX011 MgATP Solution R&D Systems Cat#B-20 Critical commercial assays Subcellular Protein Fractionation Kit Thermo Fisher Scientific Cat#78840 TriFECTa RNAi Kits Integrated DNA Technologies (IDT) N/A Experimental models: Cell lines Human: HEK293T ATCC Cat#CRL-3216 Human: HeLa Yoshida et al. 71 N/A Human: HepG2 Sakasai-Sakai et al. 72 N/A Mouse: embryonic fibroblasts Yoshida et al. 71 N/A Mouse: embryonic fibroblasts p53 − /− Dr. David J Kwiatkowski 46 N/A Experimental models: Organisms/strains B6.129P2(Cg)-Huwe1 tm1.1Mak /J The Jaxon Laboratory Hao et al. 73 Strain#029254; RRID:IMSR_JAX:029254 B6.Cg-Speer6-ps1 Tg(Alb-cre)21Mgn /J The Jaxon Laboratory Postic et al. 74 Strain#003574; RRID:IMSR_JAX:003574 Oligonucleotides shRNA targeting HUWE1-1 (H and M) 5 ′ -GCATCTGTACAGTTCCATAGA This paper N/A shRNA targeting HUWE1-2 (H and M) 5 ′ -TGCCGCAATCCAGACATAT This paper N/A (Continued on next page) 18 Cell Reports 44, 116382, October 28, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER shRNA targeting HUWE1-3 (H and M) 5 ′ -GCTCAATACTAGCCGTCTACA This paper N/A shRNA targeting HUWE1-4 (H) 5 ′ -GGGAATTTCCCACCAGGATGA This paper N/A sgRNA targeting HUWE1 (H) 5 ′ -GCTGAACTTCACCTGAATCG This paper N/A sgRNA targeting GFP 5 ′ -CTTGTCACTACTTTCGGTTA Yao et al. 20 N/A shRNA targeting CAD-1 (M) 5 ′ -GCTCAGATCCTAGTGCTAACG This paper N/A shRNA targeting CAD-2 (M) 5 ′ -GGTGGTGATGGACATCTATGA This paper N/A siRNA sequences for HUWE1, see Table S1 Integrated DNA Technologies (IDT) N/A RT-qPCR primer sequences, see Table S2 This paper N/A Recombinant DNA pLKO1-Sc (Scramble shRNA) Sarbassov et al. 75 Addgene plasmid #1864 Lenti-CRISPR v2 Sanjana et al. 76 Addgene plasmid #52961 psPAX2 Dr. Didier Trono Addgene plasmid #12260 pMD2.G Dr. Didier Trono Addgene plasmid #12259 pcDNA3-HA-Ubiquitin Kamitani et al. 77 Addgene plasmid #18712 pcDNA3-Flag-HA-CAD Ben-Sahra et al. 31 Addgene plasmid #46238 pENTR1A-HUWE1 Dr. Jeanette G Cook Addgene plasmid #37431 pENTR1A-HUWE1 C4341S Dr. Jeanette G Cook Addgene plasmid # Bioprocessing:Article Title: HUWE1 stimulates mTORC1 activity by enhancing Rheb interaction with mTORC1 and supports de novo pyrimidine synthesis. Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Mouse monoclonal anti-HA (clone 16B12) Covance Cat#MMS-101R; RRID:AB_291262 Mouse monoclonal anti-ubiquitinylated proteins (FK2) MilliporeSigma Cat#04–263; RRID:AB_612093 Alexa Fluor 594-goat polyclonal anti-rabbit IgG Thermo Fisher Scientific Cat#A11012; RRID:AB_141359 Alexa Fluor 488-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#A11029; RRID:AB_138404 Alexa Fluor 594-goat polyclonal anti-mouse IgG Thermo Fisher Scientific Cat#R37121; RRID:AB_2556549 Anti-rabbit IgG HRP-Linked Whole Ab GE Healthcare Cat#NA934; RRID:AB_772206 Anti-mouse IgG HRP-Linked Whole Ab GE Healthcare Cat#NA931; RRID:AB_772210 Bacterial and virus strains One Shot TM Stbl TM Chemically Competent E. coli Thermo Fisher Scientific Cat#C737303 Chemicals, peptides, and recombinant proteins Amino acid mixtures Sigma-Aldrich Cat#R7131 L-Glutathione reduced Sigma-Aldrich Cat#G4251 N-Ethylmaleimide (NEM) Sigma-Aldrich Cat#E3876 Saponin Sigma-Aldrich Cat#S4521 Actinomycin D Sigma-Aldrich Cat#A9415 Lipofectamine 3000 Invitrogen Cat#L3000001 Amino acid-free DMEM medium US Biological Life Sciences Cat#D9800-27 Leucine-free DMEM medium Crystalgen Cat#226-024 L-leucine RPI Cat#L22000 Dialyzed Fetal Bovine Serum Thermo Fisher Scientific Cat#26400044 Pierce TM Glutathione Agarose Thermo Fisher Scientific Cat#16100 Prolong Gold antifade reagent with DAPI Thermo Fisher Scientific Cat#P36931 Protein G Sepharose 4 Fast Flow GE Healthcare Cat#17061801 Flag peptide LifeTein Cat#LT12022 Insulin Sigma-Aldrich Cat#10908 Phosphorus-32 NEN Radiochemicals Cat#NEX011 MgATP Solution R&D Systems Cat#B-20 Critical commercial assays Subcellular Protein Fractionation Kit Thermo Fisher Scientific Cat#78840 TriFECTa RNAi Kits Integrated DNA Technologies (IDT) N/A Experimental models: Cell lines Human: HEK293T ATCC Cat#CRL-3216 Human: HeLa Yoshida et al. 71 N/A Human: HepG2 Sakasai-Sakai et al. 72 N/A Mouse: embryonic fibroblasts Yoshida et al. 71 N/A Mouse: embryonic fibroblasts p53 − /− Dr. David J Kwiatkowski 46 N/A Experimental models: Organisms/strains B6.129P2(Cg)-Huwe1 tm1.1Mak /J The Jaxon Laboratory Hao et al. 73 Strain#029254; RRID:IMSR_JAX:029254 B6.Cg-Speer6-ps1 Tg(Alb-cre)21Mgn /J The Jaxon Laboratory Postic et al. 74 Strain#003574; RRID:IMSR_JAX:003574 Oligonucleotides shRNA targeting HUWE1-1 (H and M) 5 ′ -GCATCTGTACAGTTCCATAGA This paper N/A shRNA targeting HUWE1-2 (H and M) 5 ′ -TGCCGCAATCCAGACATAT This paper N/A (Continued on next page) 18 Cell Reports 44, 116382, October 28, 2025 .. REAGENT or RESOURCE SOURCE IDENTIFIER shRNA targeting HUWE1-3 (H and M) 5 ′ -GCTCAATACTAGCCGTCTACA This paper N/A shRNA targeting HUWE1-4 (H) 5 ′ -GGGAATTTCCCACCAGGATGA This paper N/A sgRNA targeting HUWE1 (H) 5 ′ -GCTGAACTTCACCTGAATCG This paper N/A sgRNA targeting GFP 5 ′ -CTTGTCACTACTTTCGGTTA Yao et al. 20 N/A shRNA targeting CAD-1 (M) 5 ′ -GCTCAGATCCTAGTGCTAACG This paper N/A shRNA targeting CAD-2 (M) 5 ′ -GGTGGTGATGGACATCTATGA This paper N/A siRNA sequences for HUWE1, see Table S1 Integrated DNA Technologies (IDT) N/A RT-qPCR primer sequences, see Table S2 This paper N/A Recombinant DNA pLKO1-Sc (Scramble shRNA) Sarbassov et al. 75 Addgene plasmid #1864 Lenti-CRISPR v2 Sanjana et al. 76 Addgene plasmid #52961 psPAX2 Dr. Didier Trono Addgene plasmid #12260 pMD2.G Dr. Didier Trono Addgene plasmid #12259 pcDNA3-HA-Ubiquitin Kamitani et al. 77 Addgene plasmid #18712 pcDNA3-Flag-HA-CAD Ben-Sahra et al. 31 Addgene plasmid #46238 pENTR1A-HUWE1 Dr. Jeanette G Cook Addgene plasmid #37431 pENTR1A-HUWE1 C4341S Dr. Jeanette G Cook Addgene plasmid # Protein-Protein interactions:Article Title: A Novel Hap1-Tsc1 Interaction Regulates Neuronal mTORC1 Signaling and Morphogenesis in the Brain Article Snippet: .. ORFs for IP-MS were subcloned by Gateway recombination into pHAGE-N-FLAG lentiviral vector. Article Title: A Novel Hap1-Tsc1 Interaction Regulates Neuronal mTORC1 Signaling and Morphogenesis in the Brain Article Snippet: .. ORFs for IP-MS were subcloned by Gateway recombination into pHAGE-N-FLAG lentiviral vector. |
![( A ) Focused view of the TSC HR dimer. A single subunit of the TSC1 HR dimer mediates contacts to the TSC1 CC domain and the <t>TSC2</t> HRs. Two regions of the large extended loop (TSC1 IDR1) shield the exposed second binding site of the TSC1 N-terminal dimer. Top: Schematic of the TSC1 HR dimer in context of TSC. ( B ) The TSC1 HR dimer clamps onto a hydrophobic helix of the TSC1 CC domain. The second TSC1 helix is hidden for clarity [shown in (A), dark blue]. ( C ) Key contacts in the TSC1 HR dimer interface mediated by the first TSC1 extended loop (IDR1). ( D ) Focused view of the WIPI3:TSC1 interaction as resolved by cryo-EM. ( E ) The 3.17-Å crystal structure of WIPI3 in complex with the TSC1 WIR motif. Bottom: Schematic of the WIPI3 binding site in context of TSC.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_8170/pmc11578170/pmc11578170__sciadv.adr5807-f2.jpg)
