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cell lines human embryonic kidney hek 293t cells atcc crl 11268  (ATCC)


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    ATCC cell lines human embryonic kidney hek 293t cells atcc crl 11268
    Cell Lines Human Embryonic Kidney Hek 293t Cells Atcc Crl 11268, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 12205 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cell+lines+human+embryonic+kidney+hek+293t+cells+atcc+crl+11268/293T%2F17/pm41712380-245-110-118
    Average 99 stars, based on 12205 article reviews
    cell lines human embryonic kidney hek 293t cells atcc crl 11268 - by Bioz Stars, 2026-11
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    Real-time Polymerase Chain Reaction:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17

    cDNA Synthesis:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17

    Recombinant:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17

    Protease Inhibitor:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17

    Bicinchoninic Acid Protein Assay:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17

    Ubiquitin Assay:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17

    Reporter Assay:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17

    Protein Purification:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17

    Mutagenesis:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17

    shRNA:

    Article Title: Dual roles of USP39 in stabilizing PB2 and orchestrating ribonucleoprotein assembly drive H5 influenza virus replication and pathogenicity.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-STAT1 mAb Cell Signaling Technology Cat# 14995; RRID: AB_2716280 Rabbit anti-STAT2 mAb Cell Signaling Technology Cat# 72604; RRID: AB_2799824 Rabbit anti-pSTAT1 mAb Cell Signaling Technology Cat# 8826; RRID: AB_2773718 Rabbit anti-pSTAT2 mAb Cell Signaling Technology Cat# 88410; RRID: AB_2800123 Mouse anti-HA-tag mAb Cell Signaling Technology Cat# 2367; RRID: AB_10691311 Rabbit anti-K48 polyubiquitin mAb Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Mouse anti-JAK1 mAb Cell Signaling Technology Cat# 50996; RRID: AB_2716281 Rabbit anti-USP39 mAb Cell Signaling Technology Cat# 23303; RRID: AB_3720430 Rabbit anti-USP39 pAb Proteintech Cat# 23865-1-AP; RRID: AB_2879343 Mouse anti-NP mAb GeneTex Cat# GTX629633; RRID: AB_ 2888128 Rabbit anti-PB2 pAb GeneTex Cat# GTX125926; RRID: AB_11162999 Mouse anti-beta actin mAb Santa Cruz Cat# sc-47778; RRID: AB_626632 Mouse anti-His tag mAb Medical& biological laboratories Cat# D291-3; RRID: AB_10597733 Mouse anti-Myc tag mAb Medical& biological laboratories Cat# M192-3; RRID: AB_11160947 Rabbit anti-Myc tag pAb Medical& biological laboratories Cat# 562S; RRID: AB_591114 Mouse anti-FLAG M2 mAb Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse anti-PB2 pAb This paper N/A Alexa Fluor 594 Goat anti-Mouse IgG (H + L) Abcam Cat# ab150120; RRID: AB_2631447 Alexa Fluor 488 Goat anti-Rabbit IgG (H + L) Abcam Cat# ab150077; RRID: AB_2630356 Bacterial and virus strains E. coli DH5α Invitrogen Cat# 18265017 E. coli BL21 (DE3) Novagen Cat# 69450 A/chicken/Jiangsu/YB/H5N1/2015 (YB)59 Our laboratory N/A A/chicken/Jiangsu/DT0303/2016 (DT)60 Our laboratory N/A A/mallard/Huadong/S/2005 (SY)61 Our laboratory N/A A/duck/Eastern China/JY/2014 (JY)62 Our laboratory N/A A/goose/Eastern China/CZ/2013 (CZ)62 Our laboratory N/A A/Puerto Rico/8/34 (PR8)12 Our laboratory N/A VSV-GFP12 Our laboratory N/A SeV12 Our laboratory N/A Recombinant YB-PB2-K660R This paper N/A Recombinant CZ-PB2-K660R This paper N/A Chemicals, peptides, and recombinant proteins DMEM Gibco Cat# 11995065 Opti-MEM Gibco Cat# 31985062 Fetal Bovine Serum Gibco Cat# 10099141 DAPI Sigma-Aldrich Cat# D9542 Triton X-100 Beyotime Cat# P0096 Transfection Reagent BioBest Cat# B0021 NP-40 lysis buffer Beyotime Cat# P0013F RNA isolater Total RNA Extraction Reagent (TRizol) Vazyme Cat# R401-01 Protein A/G PLUS-Agarose Santa Cruz Cat# sc-2003 Dimethylsulfoxide(DMSO) Sigma-Aldrich Cat# D2650 (Continued on next page) Cell Reports 45, 117002, March 24, 2026 15 .. REAGENT or RESOURCE SOURCE IDENTIFIER MG132 Sigma-Aldrich Cat# M7449 PerfectStart Green qPCR SuperMix TransGen Cat# AQ601 TransScripte All-in-One First-Strand cDNA Synthesis Super Mix TransGen Cat# AT311 TransScript RT reagent kit Thermo Scientific Cat# 11917010 Anti-Flag M2 affinity agarose Sigma-Aldrich Cat# A2220 Recombinant human IFNβ GenScript Cat# Z03109 Ni-NTA agarose QIAGEN Cat# 30210 Protease Inhibitor Cocktail Sigma-Aldrich Cat# P2714 Critical commercial assays BCA protein assay kit Thermo Scientific Cat# 23227 SuperPico ECL Chemiluminescence Kit NCM Biotech Cat# P10300 Ubiquitylation Assay Kit Abcam Cat# ab139467 Dual-Luciferase Reporter Assay System Kit Vazyme Cat# DL101 His-tagged protein purification kit Beyotime Cat# P2247S Mut Express II Fast Mutagenesis Kit V2 Vazyme Cat# C214-02 Experimental models: Cell lines Human embryonic kidney (HEK) 293T cells ATCC CRL-11268 Human lung epithelial A549 cells ATCC CCL-185 Chicken fibroblast cell line DF-1 cells ATCC CRL-12203 Madin-Darby canine kidney (MDCK) cells ATCC CCL-34 USP39 KO HEK293T cells This paper N/A USP39 KO A549 cells This paper N/A ShUSP39 A549 cells This paper N/A Experimental models: Organisms/strains Mouse: BALB/c Laboratory Animal Center of Yangzhou University N/A Oligonucleotides Primers for real-time qPCR This paper Table S2 gRNA sequences This paper N/A shRNA sequences This paper N/A Recombinant DNA 3xFlag-PB2-YB/H5N1 This paper N/A 3xFlag-PB2-DT/H5N1 This paper N/A 3xFlag-PB2-JY/H5N8 Our laboratory N/A 3xFlag-PB2-CZ/H5N8 Our laboratory N/A 3xFlag-PB2-SY/H5N1 This paper N/A 3xFlag-PB2-PR8/H1N1 Our laboratory N/A Myc-USP39 This paper N/A Myc-USP39-C136/139A This paper N/A PB2-YB-His This paper N/A 3xFlag-PB1-YB This paper N/A 3xFlag-PA-YB This paper N/A 3xFlag-NP-YB This paper N/A pCAGGS-Fluc This paper N/A AIV MG This paper N/A ISRE-Luc Our laboratory N/A STAT1-Luc Our laboratory N/A pRL-TK Our laboratory N/A (Continued on next page) 16 Cell Reports 45, 117002, March 24, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER HA-Ub Our laboratory N/A HA-Ub-K6 Our laboratory N/A HA-Ub-K11 Our laboratory N/A HA-Ub-K27 Our laboratory N/A HA-Ub-K29 Our laboratory N/A HA-Ub-K33 Our laboratory N/A HA-Ub-K48 Our laboratory N/A HA-Ub-K63 Our laboratory N/A 3xFlag-PB2-K189R This paper N/A 3xFlag-PB2-K331R This paper N/A 3xFlag-PB2-K189/K331R This paper N/A 3xFlag-PB2-K660R This paper N/A 3xFlag-PB2(241-759aa) Our laboratory N/A 3xFlag-PB2(323-759aa) Our laboratory N/A 3xFlag-PB2(486-759aa) Our laboratory N/A 3xFlag-PB2(533-759aa) Our laboratory N/A 3xFlag-PB2(1-322aa) Our laboratory N/A 3xFlag-PB2(1-485aa Our laboratory N/A 3xFlag-PB2(1-532aa) Our laboratory N/A 3xFlag-USP39-WT This paper N/A 3xFlag-USP39(1-200aa) This paper N/A 3xFlag-USP39(201-565aa) This paper N/A PCMV-GFP This paper N/A PHW200-PB2-YB This paper N/A PHW200-PB1-YB This paper N/A PHW200-PA-YB This paper N/A PHW200-HA-YB This paper N/A PHW200-NP-YB This paper N/A PHW200-NA-YB This paper N/A PHW200-M-YB This paper N/A PHW200-NS-YB This paper N/A PHW200-PB2-CZ Our laboratory N/A PHW200-PB1-CZ Our laboratory N/A PHW200-PA-CZ Our laboratory N/A PHW200-HA-CZ Our laboratory N/A PHW200-NP-CZ Our laboratory N/A PHW200-NA-CZ Our laboratory N/A PHW200-M-CZ Our laboratory N/A PHW200-NS-CZ Our laboratory N/A PHW200-PB2-K600R-YB This paper N/A PHW200-PB2-K600R-CZ This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism V8.0.2 https://www.graphpad.com/ FlowJo V10 https://www.flowjo.com/flowjo10/ download LAS X V3.7.4 https://www.leica-microsystems.com MaxQuant V1.6.14 https://www.maxquant.org iTOL V7.3 https://itol.embl.de/itol.cgi MAFFT V7 https://mafft.cbrc.jp/alignment/software/ Cell Reports 45, 117002, March 24, 2026 17



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