Virus:Article Title: MAVS phosphorylation acts as a cellular stress sensor that modulates antiviral immunity
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-phosphoS220 MAVS Antibody Eurofins acetyl-TYGPV(phospho)SPTVS anti-Myc (9E10) Santa Cruz Technology sc-40; RRID:AB_627268 anti-HA(Y-11) Santa Cruz Technology sc-805; RRID:AB_631618 anti-p38(c-20) Santa Cruz Technology sc-535; RRID:AB_632138 anti–phospho-p38 (Thr180/Tyr182) Cell Signaling 9211; RRID:AB_331641 anti–cleaved caspase-3 (D175) 5A1E Cell Signaling 9664; RRID:AB_2070042 anti-MAVS (rodent specific) Cell Signaling 4983; RRID:AB_823566 anti–phospho-IRF3 (Ser396) 4D4G Cell Signaling 4947; RRID:AB_823547 anti-TBK1/NAK (D1B4) Cell Signaling 3504; RRID:AB_2255663 rabbit polyclonal anti-Flag Rockland Inc. 600-401-383; RRID:AB_219374 Amersham, ECL Rabbit IgG, HRP-linked whole Ab (from donkey) Cytiva NA934; RRID:AB_772206 Amersham, ECL Mouse IgG, HRP-linked whole Ab (from sheep) Cytiva NA931; RRID:AB_772210 Bacterial and virus strains Sendai virus American Type Culture Collection Cantell strain, VR-907 Vesicular stomatitis virus-green fluorescent protein (VSV-GFP) This paper Chemicals, peptides, and recombinant proteins SB202190 Med Chem Express HY-10295 SP600125 Sigma Aldrich S5567-10 MG Rotenone Sigma Aldrich R8875-1G Thapsigargin Tocris 1138 Tunicamycin LKT Labs, Inc. T8153 RNAiso TaKaRa 9108 Dulbecco’s Modified Eagle’s Medium (DMEM) Sigma Aldrich D6429 Fetal bovine serum Gibco 10437028 Penicillin-streptomycin Nacalai Tesque Inc 06168–34 Lipofectamine Transfection Reagent Invitrogen 18324020 Poly(I:C) GE Healthcare 11545784 Lipofectamine 2000 Invitrogen 11668019 GeneJuice Merck Millipore 70967 Phos-stop Roche 4906845001 Calf Intestinal Alkaline Phosphatase Takara Puromycin Sigma Aldrich P-8833 Critical commercial assays ReverTra Ace TM qPCR RT Master Mix Toyobo FSQ301 KAPA SYBR® FAST qPCR Master Mix NIPPON Genetics Co. Ltd KK4602 Dual-Luciferase® Reporter Assay System Promega E1910 Deposited data Proteomic identification of phosphorylation sites of MAVS This study https://repository.jpostdb.org/ entry/JPST003870.0 PXD064922 (Continued on next page) iScience 28, 113256, September 19, 2025 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: cell lines Mavs − /− MEFs S. Akira (Osaka University) HEK-293T Y. Gotoh (The University of Tokyo) Mavs WT MEFs This paper Mavs 2SA MEFs This paper ASK1 − /− HEK-293T This paper BHK/T7-9 cells RIKEN BRC RCB4942 Oligonucleotides Gapdh forward 5’-ATGAATACGGCTACAGCAACAGG-3 ′ Okazaki et al. 16 Gapdh reverse 5’-CTCTTGCTCAGTGTCCTTGCTG-3 ′ Okazaki et al. 16 Mouse Ifnα forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnα reverse 5’-TCACCTCCCAGGCACAGA-3 ′ Okazaki et al. 16 Mouse Ifnβ forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnβ reverse 5’-CTTTGCACCCTCCAGTAATAGC-3 ′ Okazaki et al. 16 Sendai virus P gene forward 5 ′ -GCATGGAGCCTGGCAGCTCA-3 ′ Okazaki et al. 16 Sendai virus P gene reverse 5’-CGATTCAGCGGTGGGGACCG-3 ′ Okazaki et al. 16 VSV-GFP forward 5’-GAACGGCATCAAGGTGAACT-3’ This paper VSV-GFP reverse 5’-TGCTCAGGTAGTGGTTGTCG-3’ This paper Mouse IL6 forward 5’-TGCCTTCATTTATCCCTTGAA-3 ′ This paper Mouse IL6 reverse 5’-TTACTACATTCAGCCAAAAAGCA-3 ′ This paper Mouse TNFα forward 5’-TGGCCTCCCTCTCATCAGTT-3 ′ This paper Mouse TNFα reverse 5’-GCTTGTGACTCGAATTTTGAGAAG-3 ′ This paper Mouse Mavs S220A sense primer 5’-CCTATGGACCTGTGGCTC CAACCGTTTCC-3 ′ This paper Mouse Mavs S220A anti-sense primer 5’-GGAAACGGTTGGAGCCAC AGGTCCATAGG-3 ′ This paper ASK1 gRNA forward 5’-CACCGTGCCCCTGG CATCGGTTGTC-3’ This paper ASK1 gRNA reverse 5’-AAACGACAACCGATG CCAGGGGCAC-3’ This paper Recombinant DNA PiggyBac-CAG-MAVS WT/2SA-IRES-puro This paper pCAG-hyPBase VectorBuilder pPB-CAG-empty-pgk-hph Addgene #48753 (Continued on next page) e2 iScience 28, 113256, September 19, 2025
Recombinant:Article Title: MAVS phosphorylation acts as a cellular stress sensor that modulates antiviral immunity
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-phosphoS220 MAVS Antibody Eurofins acetyl-TYGPV(phospho)SPTVS anti-Myc (9E10) Santa Cruz Technology sc-40; RRID:AB_627268 anti-HA(Y-11) Santa Cruz Technology sc-805; RRID:AB_631618 anti-p38(c-20) Santa Cruz Technology sc-535; RRID:AB_632138 anti–phospho-p38 (Thr180/Tyr182) Cell Signaling 9211; RRID:AB_331641 anti–cleaved caspase-3 (D175) 5A1E Cell Signaling 9664; RRID:AB_2070042 anti-MAVS (rodent specific) Cell Signaling 4983; RRID:AB_823566 anti–phospho-IRF3 (Ser396) 4D4G Cell Signaling 4947; RRID:AB_823547 anti-TBK1/NAK (D1B4) Cell Signaling 3504; RRID:AB_2255663 rabbit polyclonal anti-Flag Rockland Inc. 600-401-383; RRID:AB_219374 Amersham, ECL Rabbit IgG, HRP-linked whole Ab (from donkey) Cytiva NA934; RRID:AB_772206 Amersham, ECL Mouse IgG, HRP-linked whole Ab (from sheep) Cytiva NA931; RRID:AB_772210 Bacterial and virus strains Sendai virus American Type Culture Collection Cantell strain, VR-907 Vesicular stomatitis virus-green fluorescent protein (VSV-GFP) This paper Chemicals, peptides, and recombinant proteins SB202190 Med Chem Express HY-10295 SP600125 Sigma Aldrich S5567-10 MG Rotenone Sigma Aldrich R8875-1G Thapsigargin Tocris 1138 Tunicamycin LKT Labs, Inc. T8153 RNAiso TaKaRa 9108 Dulbecco’s Modified Eagle’s Medium (DMEM) Sigma Aldrich D6429 Fetal bovine serum Gibco 10437028 Penicillin-streptomycin Nacalai Tesque Inc 06168–34 Lipofectamine Transfection Reagent Invitrogen 18324020 Poly(I:C) GE Healthcare 11545784 Lipofectamine 2000 Invitrogen 11668019 GeneJuice Merck Millipore 70967 Phos-stop Roche 4906845001 Calf Intestinal Alkaline Phosphatase Takara Puromycin Sigma Aldrich P-8833 Critical commercial assays ReverTra Ace TM qPCR RT Master Mix Toyobo FSQ301 KAPA SYBR® FAST qPCR Master Mix NIPPON Genetics Co. Ltd KK4602 Dual-Luciferase® Reporter Assay System Promega E1910 Deposited data Proteomic identification of phosphorylation sites of MAVS This study https://repository.jpostdb.org/ entry/JPST003870.0 PXD064922 (Continued on next page) iScience 28, 113256, September 19, 2025 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: cell lines Mavs − /− MEFs S. Akira (Osaka University) HEK-293T Y. Gotoh (The University of Tokyo) Mavs WT MEFs This paper Mavs 2SA MEFs This paper ASK1 − /− HEK-293T This paper BHK/T7-9 cells RIKEN BRC RCB4942 Oligonucleotides Gapdh forward 5’-ATGAATACGGCTACAGCAACAGG-3 ′ Okazaki et al. 16 Gapdh reverse 5’-CTCTTGCTCAGTGTCCTTGCTG-3 ′ Okazaki et al. 16 Mouse Ifnα forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnα reverse 5’-TCACCTCCCAGGCACAGA-3 ′ Okazaki et al. 16 Mouse Ifnβ forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnβ reverse 5’-CTTTGCACCCTCCAGTAATAGC-3 ′ Okazaki et al. 16 Sendai virus P gene forward 5 ′ -GCATGGAGCCTGGCAGCTCA-3 ′ Okazaki et al. 16 Sendai virus P gene reverse 5’-CGATTCAGCGGTGGGGACCG-3 ′ Okazaki et al. 16 VSV-GFP forward 5’-GAACGGCATCAAGGTGAACT-3’ This paper VSV-GFP reverse 5’-TGCTCAGGTAGTGGTTGTCG-3’ This paper Mouse IL6 forward 5’-TGCCTTCATTTATCCCTTGAA-3 ′ This paper Mouse IL6 reverse 5’-TTACTACATTCAGCCAAAAAGCA-3 ′ This paper Mouse TNFα forward 5’-TGGCCTCCCTCTCATCAGTT-3 ′ This paper Mouse TNFα reverse 5’-GCTTGTGACTCGAATTTTGAGAAG-3 ′ This paper Mouse Mavs S220A sense primer 5’-CCTATGGACCTGTGGCTC CAACCGTTTCC-3 ′ This paper Mouse Mavs S220A anti-sense primer 5’-GGAAACGGTTGGAGCCAC AGGTCCATAGG-3 ′ This paper ASK1 gRNA forward 5’-CACCGTGCCCCTGG CATCGGTTGTC-3’ This paper ASK1 gRNA reverse 5’-AAACGACAACCGATG CCAGGGGCAC-3’ This paper Recombinant DNA PiggyBac-CAG-MAVS WT/2SA-IRES-puro This paper pCAG-hyPBase VectorBuilder pPB-CAG-empty-pgk-hph Addgene #48753 (Continued on next page) e2 iScience 28, 113256, September 19, 2025
Modification:Article Title: MAVS phosphorylation acts as a cellular stress sensor that modulates antiviral immunity
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-phosphoS220 MAVS Antibody Eurofins acetyl-TYGPV(phospho)SPTVS anti-Myc (9E10) Santa Cruz Technology sc-40; RRID:AB_627268 anti-HA(Y-11) Santa Cruz Technology sc-805; RRID:AB_631618 anti-p38(c-20) Santa Cruz Technology sc-535; RRID:AB_632138 anti–phospho-p38 (Thr180/Tyr182) Cell Signaling 9211; RRID:AB_331641 anti–cleaved caspase-3 (D175) 5A1E Cell Signaling 9664; RRID:AB_2070042 anti-MAVS (rodent specific) Cell Signaling 4983; RRID:AB_823566 anti–phospho-IRF3 (Ser396) 4D4G Cell Signaling 4947; RRID:AB_823547 anti-TBK1/NAK (D1B4) Cell Signaling 3504; RRID:AB_2255663 rabbit polyclonal anti-Flag Rockland Inc. 600-401-383; RRID:AB_219374 Amersham, ECL Rabbit IgG, HRP-linked whole Ab (from donkey) Cytiva NA934; RRID:AB_772206 Amersham, ECL Mouse IgG, HRP-linked whole Ab (from sheep) Cytiva NA931; RRID:AB_772210 Bacterial and virus strains Sendai virus American Type Culture Collection Cantell strain, VR-907 Vesicular stomatitis virus-green fluorescent protein (VSV-GFP) This paper Chemicals, peptides, and recombinant proteins SB202190 Med Chem Express HY-10295 SP600125 Sigma Aldrich S5567-10 MG Rotenone Sigma Aldrich R8875-1G Thapsigargin Tocris 1138 Tunicamycin LKT Labs, Inc. T8153 RNAiso TaKaRa 9108 Dulbecco’s Modified Eagle’s Medium (DMEM) Sigma Aldrich D6429 Fetal bovine serum Gibco 10437028 Penicillin-streptomycin Nacalai Tesque Inc 06168–34 Lipofectamine Transfection Reagent Invitrogen 18324020 Poly(I:C) GE Healthcare 11545784 Lipofectamine 2000 Invitrogen 11668019 GeneJuice Merck Millipore 70967 Phos-stop Roche 4906845001 Calf Intestinal Alkaline Phosphatase Takara Puromycin Sigma Aldrich P-8833 Critical commercial assays ReverTra Ace TM qPCR RT Master Mix Toyobo FSQ301 KAPA SYBR® FAST qPCR Master Mix NIPPON Genetics Co. Ltd KK4602 Dual-Luciferase® Reporter Assay System Promega E1910 Deposited data Proteomic identification of phosphorylation sites of MAVS This study https://repository.jpostdb.org/ entry/JPST003870.0 PXD064922 (Continued on next page) iScience 28, 113256, September 19, 2025 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: cell lines Mavs − /− MEFs S. Akira (Osaka University) HEK-293T Y. Gotoh (The University of Tokyo) Mavs WT MEFs This paper Mavs 2SA MEFs This paper ASK1 − /− HEK-293T This paper BHK/T7-9 cells RIKEN BRC RCB4942 Oligonucleotides Gapdh forward 5’-ATGAATACGGCTACAGCAACAGG-3 ′ Okazaki et al. 16 Gapdh reverse 5’-CTCTTGCTCAGTGTCCTTGCTG-3 ′ Okazaki et al. 16 Mouse Ifnα forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnα reverse 5’-TCACCTCCCAGGCACAGA-3 ′ Okazaki et al. 16 Mouse Ifnβ forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnβ reverse 5’-CTTTGCACCCTCCAGTAATAGC-3 ′ Okazaki et al. 16 Sendai virus P gene forward 5 ′ -GCATGGAGCCTGGCAGCTCA-3 ′ Okazaki et al. 16 Sendai virus P gene reverse 5’-CGATTCAGCGGTGGGGACCG-3 ′ Okazaki et al. 16 VSV-GFP forward 5’-GAACGGCATCAAGGTGAACT-3’ This paper VSV-GFP reverse 5’-TGCTCAGGTAGTGGTTGTCG-3’ This paper Mouse IL6 forward 5’-TGCCTTCATTTATCCCTTGAA-3 ′ This paper Mouse IL6 reverse 5’-TTACTACATTCAGCCAAAAAGCA-3 ′ This paper Mouse TNFα forward 5’-TGGCCTCCCTCTCATCAGTT-3 ′ This paper Mouse TNFα reverse 5’-GCTTGTGACTCGAATTTTGAGAAG-3 ′ This paper Mouse Mavs S220A sense primer 5’-CCTATGGACCTGTGGCTC CAACCGTTTCC-3 ′ This paper Mouse Mavs S220A anti-sense primer 5’-GGAAACGGTTGGAGCCAC AGGTCCATAGG-3 ′ This paper ASK1 gRNA forward 5’-CACCGTGCCCCTGG CATCGGTTGTC-3’ This paper ASK1 gRNA reverse 5’-AAACGACAACCGATG CCAGGGGCAC-3’ This paper Recombinant DNA PiggyBac-CAG-MAVS WT/2SA-IRES-puro This paper pCAG-hyPBase VectorBuilder pPB-CAG-empty-pgk-hph Addgene #48753 (Continued on next page) e2 iScience 28, 113256, September 19, 2025
Transfection:Article Title: MAVS phosphorylation acts as a cellular stress sensor that modulates antiviral immunity
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-phosphoS220 MAVS Antibody Eurofins acetyl-TYGPV(phospho)SPTVS anti-Myc (9E10) Santa Cruz Technology sc-40; RRID:AB_627268 anti-HA(Y-11) Santa Cruz Technology sc-805; RRID:AB_631618 anti-p38(c-20) Santa Cruz Technology sc-535; RRID:AB_632138 anti–phospho-p38 (Thr180/Tyr182) Cell Signaling 9211; RRID:AB_331641 anti–cleaved caspase-3 (D175) 5A1E Cell Signaling 9664; RRID:AB_2070042 anti-MAVS (rodent specific) Cell Signaling 4983; RRID:AB_823566 anti–phospho-IRF3 (Ser396) 4D4G Cell Signaling 4947; RRID:AB_823547 anti-TBK1/NAK (D1B4) Cell Signaling 3504; RRID:AB_2255663 rabbit polyclonal anti-Flag Rockland Inc. 600-401-383; RRID:AB_219374 Amersham, ECL Rabbit IgG, HRP-linked whole Ab (from donkey) Cytiva NA934; RRID:AB_772206 Amersham, ECL Mouse IgG, HRP-linked whole Ab (from sheep) Cytiva NA931; RRID:AB_772210 Bacterial and virus strains Sendai virus American Type Culture Collection Cantell strain, VR-907 Vesicular stomatitis virus-green fluorescent protein (VSV-GFP) This paper Chemicals, peptides, and recombinant proteins SB202190 Med Chem Express HY-10295 SP600125 Sigma Aldrich S5567-10 MG Rotenone Sigma Aldrich R8875-1G Thapsigargin Tocris 1138 Tunicamycin LKT Labs, Inc. T8153 RNAiso TaKaRa 9108 Dulbecco’s Modified Eagle’s Medium (DMEM) Sigma Aldrich D6429 Fetal bovine serum Gibco 10437028 Penicillin-streptomycin Nacalai Tesque Inc 06168–34 Lipofectamine Transfection Reagent Invitrogen 18324020 Poly(I:C) GE Healthcare 11545784 Lipofectamine 2000 Invitrogen 11668019 GeneJuice Merck Millipore 70967 Phos-stop Roche 4906845001 Calf Intestinal Alkaline Phosphatase Takara Puromycin Sigma Aldrich P-8833 Critical commercial assays ReverTra Ace TM qPCR RT Master Mix Toyobo FSQ301 KAPA SYBR® FAST qPCR Master Mix NIPPON Genetics Co. Ltd KK4602 Dual-Luciferase® Reporter Assay System Promega E1910 Deposited data Proteomic identification of phosphorylation sites of MAVS This study https://repository.jpostdb.org/ entry/JPST003870.0 PXD064922 (Continued on next page) iScience 28, 113256, September 19, 2025 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: cell lines Mavs − /− MEFs S. Akira (Osaka University) HEK-293T Y. Gotoh (The University of Tokyo) Mavs WT MEFs This paper Mavs 2SA MEFs This paper ASK1 − /− HEK-293T This paper BHK/T7-9 cells RIKEN BRC RCB4942 Oligonucleotides Gapdh forward 5’-ATGAATACGGCTACAGCAACAGG-3 ′ Okazaki et al. 16 Gapdh reverse 5’-CTCTTGCTCAGTGTCCTTGCTG-3 ′ Okazaki et al. 16 Mouse Ifnα forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnα reverse 5’-TCACCTCCCAGGCACAGA-3 ′ Okazaki et al. 16 Mouse Ifnβ forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnβ reverse 5’-CTTTGCACCCTCCAGTAATAGC-3 ′ Okazaki et al. 16 Sendai virus P gene forward 5 ′ -GCATGGAGCCTGGCAGCTCA-3 ′ Okazaki et al. 16 Sendai virus P gene reverse 5’-CGATTCAGCGGTGGGGACCG-3 ′ Okazaki et al. 16 VSV-GFP forward 5’-GAACGGCATCAAGGTGAACT-3’ This paper VSV-GFP reverse 5’-TGCTCAGGTAGTGGTTGTCG-3’ This paper Mouse IL6 forward 5’-TGCCTTCATTTATCCCTTGAA-3 ′ This paper Mouse IL6 reverse 5’-TTACTACATTCAGCCAAAAAGCA-3 ′ This paper Mouse TNFα forward 5’-TGGCCTCCCTCTCATCAGTT-3 ′ This paper Mouse TNFα reverse 5’-GCTTGTGACTCGAATTTTGAGAAG-3 ′ This paper Mouse Mavs S220A sense primer 5’-CCTATGGACCTGTGGCTC CAACCGTTTCC-3 ′ This paper Mouse Mavs S220A anti-sense primer 5’-GGAAACGGTTGGAGCCAC AGGTCCATAGG-3 ′ This paper ASK1 gRNA forward 5’-CACCGTGCCCCTGG CATCGGTTGTC-3’ This paper ASK1 gRNA reverse 5’-AAACGACAACCGATG CCAGGGGCAC-3’ This paper Recombinant DNA PiggyBac-CAG-MAVS WT/2SA-IRES-puro This paper pCAG-hyPBase VectorBuilder pPB-CAG-empty-pgk-hph Addgene #48753 (Continued on next page) e2 iScience 28, 113256, September 19, 2025
Article Title: LncRNA HITT inhibits autophagy by attenuating ATG12-ATG5-ATG16L1 complex formation.
Article Snippet: Dysregulated autophagy has been implicated in the pathogenesis of numerous diseases, including cancer.. Despite extensive research on the underlying mechanisms of autophagy, the involvement of long non-coding RNAs (lncRNAs) remains poorly understood.. Here, we demonstrate that a previously identified lncRNA, HITT (HIF-1α inhibitor at the translation level), is closely associated with biological processes such as autophagy through unbiased bioinformatic analysis.
Real-time Polymerase Chain Reaction:Article Title: MAVS phosphorylation acts as a cellular stress sensor that modulates antiviral immunity
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-phosphoS220 MAVS Antibody Eurofins acetyl-TYGPV(phospho)SPTVS anti-Myc (9E10) Santa Cruz Technology sc-40; RRID:AB_627268 anti-HA(Y-11) Santa Cruz Technology sc-805; RRID:AB_631618 anti-p38(c-20) Santa Cruz Technology sc-535; RRID:AB_632138 anti–phospho-p38 (Thr180/Tyr182) Cell Signaling 9211; RRID:AB_331641 anti–cleaved caspase-3 (D175) 5A1E Cell Signaling 9664; RRID:AB_2070042 anti-MAVS (rodent specific) Cell Signaling 4983; RRID:AB_823566 anti–phospho-IRF3 (Ser396) 4D4G Cell Signaling 4947; RRID:AB_823547 anti-TBK1/NAK (D1B4) Cell Signaling 3504; RRID:AB_2255663 rabbit polyclonal anti-Flag Rockland Inc. 600-401-383; RRID:AB_219374 Amersham, ECL Rabbit IgG, HRP-linked whole Ab (from donkey) Cytiva NA934; RRID:AB_772206 Amersham, ECL Mouse IgG, HRP-linked whole Ab (from sheep) Cytiva NA931; RRID:AB_772210 Bacterial and virus strains Sendai virus American Type Culture Collection Cantell strain, VR-907 Vesicular stomatitis virus-green fluorescent protein (VSV-GFP) This paper Chemicals, peptides, and recombinant proteins SB202190 Med Chem Express HY-10295 SP600125 Sigma Aldrich S5567-10 MG Rotenone Sigma Aldrich R8875-1G Thapsigargin Tocris 1138 Tunicamycin LKT Labs, Inc. T8153 RNAiso TaKaRa 9108 Dulbecco’s Modified Eagle’s Medium (DMEM) Sigma Aldrich D6429 Fetal bovine serum Gibco 10437028 Penicillin-streptomycin Nacalai Tesque Inc 06168–34 Lipofectamine Transfection Reagent Invitrogen 18324020 Poly(I:C) GE Healthcare 11545784 Lipofectamine 2000 Invitrogen 11668019 GeneJuice Merck Millipore 70967 Phos-stop Roche 4906845001 Calf Intestinal Alkaline Phosphatase Takara Puromycin Sigma Aldrich P-8833 Critical commercial assays ReverTra Ace TM qPCR RT Master Mix Toyobo FSQ301 KAPA SYBR® FAST qPCR Master Mix NIPPON Genetics Co. Ltd KK4602 Dual-Luciferase® Reporter Assay System Promega E1910 Deposited data Proteomic identification of phosphorylation sites of MAVS This study https://repository.jpostdb.org/ entry/JPST003870.0 PXD064922 (Continued on next page) iScience 28, 113256, September 19, 2025 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: cell lines Mavs − /− MEFs S. Akira (Osaka University) HEK-293T Y. Gotoh (The University of Tokyo) Mavs WT MEFs This paper Mavs 2SA MEFs This paper ASK1 − /− HEK-293T This paper BHK/T7-9 cells RIKEN BRC RCB4942 Oligonucleotides Gapdh forward 5’-ATGAATACGGCTACAGCAACAGG-3 ′ Okazaki et al. 16 Gapdh reverse 5’-CTCTTGCTCAGTGTCCTTGCTG-3 ′ Okazaki et al. 16 Mouse Ifnα forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnα reverse 5’-TCACCTCCCAGGCACAGA-3 ′ Okazaki et al. 16 Mouse Ifnβ forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnβ reverse 5’-CTTTGCACCCTCCAGTAATAGC-3 ′ Okazaki et al. 16 Sendai virus P gene forward 5 ′ -GCATGGAGCCTGGCAGCTCA-3 ′ Okazaki et al. 16 Sendai virus P gene reverse 5’-CGATTCAGCGGTGGGGACCG-3 ′ Okazaki et al. 16 VSV-GFP forward 5’-GAACGGCATCAAGGTGAACT-3’ This paper VSV-GFP reverse 5’-TGCTCAGGTAGTGGTTGTCG-3’ This paper Mouse IL6 forward 5’-TGCCTTCATTTATCCCTTGAA-3 ′ This paper Mouse IL6 reverse 5’-TTACTACATTCAGCCAAAAAGCA-3 ′ This paper Mouse TNFα forward 5’-TGGCCTCCCTCTCATCAGTT-3 ′ This paper Mouse TNFα reverse 5’-GCTTGTGACTCGAATTTTGAGAAG-3 ′ This paper Mouse Mavs S220A sense primer 5’-CCTATGGACCTGTGGCTC CAACCGTTTCC-3 ′ This paper Mouse Mavs S220A anti-sense primer 5’-GGAAACGGTTGGAGCCAC AGGTCCATAGG-3 ′ This paper ASK1 gRNA forward 5’-CACCGTGCCCCTGG CATCGGTTGTC-3’ This paper ASK1 gRNA reverse 5’-AAACGACAACCGATG CCAGGGGCAC-3’ This paper Recombinant DNA PiggyBac-CAG-MAVS WT/2SA-IRES-puro This paper pCAG-hyPBase VectorBuilder pPB-CAG-empty-pgk-hph Addgene #48753 (Continued on next page) e2 iScience 28, 113256, September 19, 2025
Reporter Assay:Article Title: MAVS phosphorylation acts as a cellular stress sensor that modulates antiviral immunity
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-phosphoS220 MAVS Antibody Eurofins acetyl-TYGPV(phospho)SPTVS anti-Myc (9E10) Santa Cruz Technology sc-40; RRID:AB_627268 anti-HA(Y-11) Santa Cruz Technology sc-805; RRID:AB_631618 anti-p38(c-20) Santa Cruz Technology sc-535; RRID:AB_632138 anti–phospho-p38 (Thr180/Tyr182) Cell Signaling 9211; RRID:AB_331641 anti–cleaved caspase-3 (D175) 5A1E Cell Signaling 9664; RRID:AB_2070042 anti-MAVS (rodent specific) Cell Signaling 4983; RRID:AB_823566 anti–phospho-IRF3 (Ser396) 4D4G Cell Signaling 4947; RRID:AB_823547 anti-TBK1/NAK (D1B4) Cell Signaling 3504; RRID:AB_2255663 rabbit polyclonal anti-Flag Rockland Inc. 600-401-383; RRID:AB_219374 Amersham, ECL Rabbit IgG, HRP-linked whole Ab (from donkey) Cytiva NA934; RRID:AB_772206 Amersham, ECL Mouse IgG, HRP-linked whole Ab (from sheep) Cytiva NA931; RRID:AB_772210 Bacterial and virus strains Sendai virus American Type Culture Collection Cantell strain, VR-907 Vesicular stomatitis virus-green fluorescent protein (VSV-GFP) This paper Chemicals, peptides, and recombinant proteins SB202190 Med Chem Express HY-10295 SP600125 Sigma Aldrich S5567-10 MG Rotenone Sigma Aldrich R8875-1G Thapsigargin Tocris 1138 Tunicamycin LKT Labs, Inc. T8153 RNAiso TaKaRa 9108 Dulbecco’s Modified Eagle’s Medium (DMEM) Sigma Aldrich D6429 Fetal bovine serum Gibco 10437028 Penicillin-streptomycin Nacalai Tesque Inc 06168–34 Lipofectamine Transfection Reagent Invitrogen 18324020 Poly(I:C) GE Healthcare 11545784 Lipofectamine 2000 Invitrogen 11668019 GeneJuice Merck Millipore 70967 Phos-stop Roche 4906845001 Calf Intestinal Alkaline Phosphatase Takara Puromycin Sigma Aldrich P-8833 Critical commercial assays ReverTra Ace TM qPCR RT Master Mix Toyobo FSQ301 KAPA SYBR® FAST qPCR Master Mix NIPPON Genetics Co. Ltd KK4602 Dual-Luciferase® Reporter Assay System Promega E1910 Deposited data Proteomic identification of phosphorylation sites of MAVS This study https://repository.jpostdb.org/ entry/JPST003870.0 PXD064922 (Continued on next page) iScience 28, 113256, September 19, 2025 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: cell lines Mavs − /− MEFs S. Akira (Osaka University) HEK-293T Y. Gotoh (The University of Tokyo) Mavs WT MEFs This paper Mavs 2SA MEFs This paper ASK1 − /− HEK-293T This paper BHK/T7-9 cells RIKEN BRC RCB4942 Oligonucleotides Gapdh forward 5’-ATGAATACGGCTACAGCAACAGG-3 ′ Okazaki et al. 16 Gapdh reverse 5’-CTCTTGCTCAGTGTCCTTGCTG-3 ′ Okazaki et al. 16 Mouse Ifnα forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnα reverse 5’-TCACCTCCCAGGCACAGA-3 ′ Okazaki et al. 16 Mouse Ifnβ forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnβ reverse 5’-CTTTGCACCCTCCAGTAATAGC-3 ′ Okazaki et al. 16 Sendai virus P gene forward 5 ′ -GCATGGAGCCTGGCAGCTCA-3 ′ Okazaki et al. 16 Sendai virus P gene reverse 5’-CGATTCAGCGGTGGGGACCG-3 ′ Okazaki et al. 16 VSV-GFP forward 5’-GAACGGCATCAAGGTGAACT-3’ This paper VSV-GFP reverse 5’-TGCTCAGGTAGTGGTTGTCG-3’ This paper Mouse IL6 forward 5’-TGCCTTCATTTATCCCTTGAA-3 ′ This paper Mouse IL6 reverse 5’-TTACTACATTCAGCCAAAAAGCA-3 ′ This paper Mouse TNFα forward 5’-TGGCCTCCCTCTCATCAGTT-3 ′ This paper Mouse TNFα reverse 5’-GCTTGTGACTCGAATTTTGAGAAG-3 ′ This paper Mouse Mavs S220A sense primer 5’-CCTATGGACCTGTGGCTC CAACCGTTTCC-3 ′ This paper Mouse Mavs S220A anti-sense primer 5’-GGAAACGGTTGGAGCCAC AGGTCCATAGG-3 ′ This paper ASK1 gRNA forward 5’-CACCGTGCCCCTGG CATCGGTTGTC-3’ This paper ASK1 gRNA reverse 5’-AAACGACAACCGATG CCAGGGGCAC-3’ This paper Recombinant DNA PiggyBac-CAG-MAVS WT/2SA-IRES-puro This paper pCAG-hyPBase VectorBuilder pPB-CAG-empty-pgk-hph Addgene #48753 (Continued on next page) e2 iScience 28, 113256, September 19, 2025
Phospho-proteomics:Article Title: MAVS phosphorylation acts as a cellular stress sensor that modulates antiviral immunity
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-phosphoS220 MAVS Antibody Eurofins acetyl-TYGPV(phospho)SPTVS anti-Myc (9E10) Santa Cruz Technology sc-40; RRID:AB_627268 anti-HA(Y-11) Santa Cruz Technology sc-805; RRID:AB_631618 anti-p38(c-20) Santa Cruz Technology sc-535; RRID:AB_632138 anti–phospho-p38 (Thr180/Tyr182) Cell Signaling 9211; RRID:AB_331641 anti–cleaved caspase-3 (D175) 5A1E Cell Signaling 9664; RRID:AB_2070042 anti-MAVS (rodent specific) Cell Signaling 4983; RRID:AB_823566 anti–phospho-IRF3 (Ser396) 4D4G Cell Signaling 4947; RRID:AB_823547 anti-TBK1/NAK (D1B4) Cell Signaling 3504; RRID:AB_2255663 rabbit polyclonal anti-Flag Rockland Inc. 600-401-383; RRID:AB_219374 Amersham, ECL Rabbit IgG, HRP-linked whole Ab (from donkey) Cytiva NA934; RRID:AB_772206 Amersham, ECL Mouse IgG, HRP-linked whole Ab (from sheep) Cytiva NA931; RRID:AB_772210 Bacterial and virus strains Sendai virus American Type Culture Collection Cantell strain, VR-907 Vesicular stomatitis virus-green fluorescent protein (VSV-GFP) This paper Chemicals, peptides, and recombinant proteins SB202190 Med Chem Express HY-10295 SP600125 Sigma Aldrich S5567-10 MG Rotenone Sigma Aldrich R8875-1G Thapsigargin Tocris 1138 Tunicamycin LKT Labs, Inc. T8153 RNAiso TaKaRa 9108 Dulbecco’s Modified Eagle’s Medium (DMEM) Sigma Aldrich D6429 Fetal bovine serum Gibco 10437028 Penicillin-streptomycin Nacalai Tesque Inc 06168–34 Lipofectamine Transfection Reagent Invitrogen 18324020 Poly(I:C) GE Healthcare 11545784 Lipofectamine 2000 Invitrogen 11668019 GeneJuice Merck Millipore 70967 Phos-stop Roche 4906845001 Calf Intestinal Alkaline Phosphatase Takara Puromycin Sigma Aldrich P-8833 Critical commercial assays ReverTra Ace TM qPCR RT Master Mix Toyobo FSQ301 KAPA SYBR® FAST qPCR Master Mix NIPPON Genetics Co. Ltd KK4602 Dual-Luciferase® Reporter Assay System Promega E1910 Deposited data Proteomic identification of phosphorylation sites of MAVS This study https://repository.jpostdb.org/ entry/JPST003870.0 PXD064922 (Continued on next page) iScience 28, 113256, September 19, 2025 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: cell lines Mavs − /− MEFs S. Akira (Osaka University) HEK-293T Y. Gotoh (The University of Tokyo) Mavs WT MEFs This paper Mavs 2SA MEFs This paper ASK1 − /− HEK-293T This paper BHK/T7-9 cells RIKEN BRC RCB4942 Oligonucleotides Gapdh forward 5’-ATGAATACGGCTACAGCAACAGG-3 ′ Okazaki et al. 16 Gapdh reverse 5’-CTCTTGCTCAGTGTCCTTGCTG-3 ′ Okazaki et al. 16 Mouse Ifnα forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnα reverse 5’-TCACCTCCCAGGCACAGA-3 ′ Okazaki et al. 16 Mouse Ifnβ forward 5’-CCTGTGTGATGCAGGAACC-3 ′ Okazaki et al. 16 Mouse Ifnβ reverse 5’-CTTTGCACCCTCCAGTAATAGC-3 ′ Okazaki et al. 16 Sendai virus P gene forward 5 ′ -GCATGGAGCCTGGCAGCTCA-3 ′ Okazaki et al. 16 Sendai virus P gene reverse 5’-CGATTCAGCGGTGGGGACCG-3 ′ Okazaki et al. 16 VSV-GFP forward 5’-GAACGGCATCAAGGTGAACT-3’ This paper VSV-GFP reverse 5’-TGCTCAGGTAGTGGTTGTCG-3’ This paper Mouse IL6 forward 5’-TGCCTTCATTTATCCCTTGAA-3 ′ This paper Mouse IL6 reverse 5’-TTACTACATTCAGCCAAAAAGCA-3 ′ This paper Mouse TNFα forward 5’-TGGCCTCCCTCTCATCAGTT-3 ′ This paper Mouse TNFα reverse 5’-GCTTGTGACTCGAATTTTGAGAAG-3 ′ This paper Mouse Mavs S220A sense primer 5’-CCTATGGACCTGTGGCTC CAACCGTTTCC-3 ′ This paper Mouse Mavs S220A anti-sense primer 5’-GGAAACGGTTGGAGCCAC AGGTCCATAGG-3 ′ This paper ASK1 gRNA forward 5’-CACCGTGCCCCTGG CATCGGTTGTC-3’ This paper ASK1 gRNA reverse 5’-AAACGACAACCGATG CCAGGGGCAC-3’ This paper Recombinant DNA PiggyBac-CAG-MAVS WT/2SA-IRES-puro This paper pCAG-hyPBase VectorBuilder pPB-CAG-empty-pgk-hph Addgene #48753 (Continued on next page) e2 iScience 28, 113256, September 19, 2025
Plasmid Preparation:Article Title: LncRNA HITT inhibits autophagy by attenuating ATG12-ATG5-ATG16L1 complex formation.
Article Snippet: Dysregulated autophagy has been implicated in the pathogenesis of numerous diseases, including cancer.. Despite extensive research on the underlying mechanisms of autophagy, the involvement of long non-coding RNAs (lncRNAs) remains poorly understood.. Here, we demonstrate that a previously identified lncRNA, HITT (HIF-1α inhibitor at the translation level), is closely associated with biological processes such as autophagy through unbiased bioinformatic analysis.
Fluorescence:
Western Blot:
Expressing:
Binding Assay:Article Title: Targeted degradation of Drosophila FOG homolog U-shaped mimics macrophage transdifferentiation in S2 cells
Article Snippet: .. In order to stabilize the F-box protein [FKBP(L106P)-Nslmb-vhhGFP4 fusion protein] [ ] and to achieve binding of the GFP-tagged protein, the small molecule inhibitor Shield-1 (HY-112210, MedChemExpress) was added to the cells with a final concentration of 5 μM. .. For sufficient induction of the GFP-Degron system CuSO 4 (P023.1, Roth) is added to cells to a final concentration of 100 μM.
Concentration Assay:Article Title: Targeted degradation of Drosophila FOG homolog U-shaped mimics macrophage transdifferentiation in S2 cells
Article Snippet: .. In order to stabilize the F-box protein [FKBP(L106P)-Nslmb-vhhGFP4 fusion protein] [ ] and to achieve binding of the GFP-tagged protein, the small molecule inhibitor Shield-1 (HY-112210, MedChemExpress) was added to the cells with a final concentration of 5 μM. .. For sufficient induction of the GFP-Degron system CuSO 4 (P023.1, Roth) is added to cells to a final concentration of 100 μM.
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