mouse anti nf κb Search Results


irf4  (Miltenyi Biotec)
94
Miltenyi Biotec irf4
Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, <t>IRF4,</t> and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.
Irf4, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/IRF-4+Antibody%2C+anti-human%2Fmouse%2C+REAfinity/bio_rxiv__64898__2026__01__19__700410-173-12-6
Average 94 stars, based on 1 article reviews
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94/100 stars
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anti il 31 pe  (Miltenyi Biotec)
92
Miltenyi Biotec anti il 31 pe
Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, <t>IRF4,</t> and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.
Anti Il 31 Pe, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/Oct-2+Antibody%2C+anti-human%2Fmouse%2C+REAfinity/pmc10619474__ADVS___8___2004683___s001-91-4-1
Average 92 stars, based on 1 article reviews
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92/100 stars
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rela nf κb p65  (R&D Systems)
94
R&D Systems rela nf κb p65
Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, <t>IRF4,</t> and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.
Rela Nf κb P65, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/Human%2FMouse%2FRat+RelA%2FNF+kappa+B+p65+Antibody/pm28620671-48-8-11
Average 94 stars, based on 1 article reviews
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94/100 stars
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anti p65 nf κb  (R&D Systems)
93
R&D Systems anti p65 nf κb
Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, <t>IRF4,</t> and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.
Anti P65 Nf κb, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/Human%2FMouse%2FRat+RelA%2FNF+kappa+B+p65+Antibody/pm37583906-49-3-14
Average 93 stars, based on 1 article reviews
anti p65 nf κb - by Bioz Stars, 2026-10
93/100 stars
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oct3 4 apc  (Miltenyi Biotec)
94
Miltenyi Biotec oct3 4 apc
Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, <t>IRF4,</t> and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.
Oct3 4 Apc, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/Oct3%2F4+Isoform+A+Antibody%2C+anti-human%2Fmouse%2C+REAfinity/pm38771392-363-45-47
Average 94 stars, based on 1 article reviews
oct3 4 apc - by Bioz Stars, 2026-10
94/100 stars
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b v co  (Miltenyi Biotec)
94
Miltenyi Biotec b v co
Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, <t>IRF4,</t> and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.
B V Co, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/Neurofilament+Antibody%2C+anti-human%2Fmouse%2C+REAfinity/pmc12796729-24-8-6
Average 94 stars, based on 1 article reviews
b v co - by Bioz Stars, 2026-10
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7074 anti total p65 r d systems  (R&D Systems)
92
R&D Systems 7074 anti total p65 r d systems
Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, <t>IRF4,</t> and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.
7074 Anti Total P65 R D Systems, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/Human%2FMouse%2FRat+RelA%2FNF+kappa+B+p65+Antibody/pmc10986120__13046_2024_3019_MOESM2_ESM-35-2-5
Average 92 stars, based on 1 article reviews
7074 anti total p65 r d systems - by Bioz Stars, 2026-10
92/100 stars
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mouse epha2 rea579  (Miltenyi Biotec)
94
Miltenyi Biotec mouse epha2 rea579
Figure 1 Vector construction, validation and assessment of <t>EphA2</t> expression. (A) Schematic of the C170 and C172 EphA2 expression viruses. (B) Southern blot confirming the anticipated NcoI fragments in our new recombinants (1.2 and 3.0 kb, C170; 2.3 and 3.0 kb, C172), (C) Immunofluorescent imaging shows different cellular distributions of the full length and secreted forms of the EphA2. (D) Western blots of CT2A infected cells and supernatants show that C170-expressed EphA2 remains cell associated, whereas C172 expressing the extracellular form of EphA2 secretes the protein into the supernatant. TAA: tumor- associated antigen. UTR: untranslated region. TGN: Trans Golgi Network. DAPI: 4′,6-diamidino-2-phenylindole
Mouse Epha2 Rea579, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/EphA2+Antibody%2C+anti-mouse%2C+REAfinity/pm34599026-83-60-64
Average 94 stars, based on 1 article reviews
mouse epha2 rea579 - by Bioz Stars, 2026-10
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axons  (Rockland Immunochemicals)
85
Rockland Immunochemicals axons
Figure 1 Vector construction, validation and assessment of <t>EphA2</t> expression. (A) Schematic of the C170 and C172 EphA2 expression viruses. (B) Southern blot confirming the anticipated NcoI fragments in our new recombinants (1.2 and 3.0 kb, C170; 2.3 and 3.0 kb, C172), (C) Immunofluorescent imaging shows different cellular distributions of the full length and secreted forms of the EphA2. (D) Western blots of CT2A infected cells and supernatants show that C170-expressed EphA2 remains cell associated, whereas C172 expressing the extracellular form of EphA2 secretes the protein into the supernatant. TAA: tumor- associated antigen. UTR: untranslated region. TGN: Trans Golgi Network. DAPI: 4′,6-diamidino-2-phenylindole
Axons, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/Neurofilament+H+Antibody/pmc06605374-328-17-23
Average 85 stars, based on 1 article reviews
axons - by Bioz Stars, 2026-10
85/100 stars
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mouse anti nf antibody  (Bio-Rad)
86
Bio-Rad mouse anti nf antibody
Figure 1 Vector construction, validation and assessment of <t>EphA2</t> expression. (A) Schematic of the C170 and C172 EphA2 expression viruses. (B) Southern blot confirming the anticipated NcoI fragments in our new recombinants (1.2 and 3.0 kb, C170; 2.3 and 3.0 kb, C172), (C) Immunofluorescent imaging shows different cellular distributions of the full length and secreted forms of the EphA2. (D) Western blots of CT2A infected cells and supernatants show that C170-expressed EphA2 remains cell associated, whereas C172 expressing the extracellular form of EphA2 secretes the protein into the supernatant. TAA: tumor- associated antigen. UTR: untranslated region. TGN: Trans Golgi Network. DAPI: 4′,6-diamidino-2-phenylindole
Mouse Anti Nf Antibody, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/Mouse+anti+NF+Kappa+B+Inhibitor+Alpha/10__1523_slash_jneurosci__4345___10__2011-116-17-24
Average 86 stars, based on 1 article reviews
mouse anti nf antibody - by Bioz Stars, 2026-10
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purified 4 mouse anti-nf-atc3 monoclonal antibody  (Becton Dickinson)
90
Becton Dickinson purified 4 mouse anti-nf-atc3 monoclonal antibody
Figure 1 Vector construction, validation and assessment of <t>EphA2</t> expression. (A) Schematic of the C170 and C172 EphA2 expression viruses. (B) Southern blot confirming the anticipated NcoI fragments in our new recombinants (1.2 and 3.0 kb, C170; 2.3 and 3.0 kb, C172), (C) Immunofluorescent imaging shows different cellular distributions of the full length and secreted forms of the EphA2. (D) Western blots of CT2A infected cells and supernatants show that C170-expressed EphA2 remains cell associated, whereas C172 expressing the extracellular form of EphA2 secretes the protein into the supernatant. TAA: tumor- associated antigen. UTR: untranslated region. TGN: Trans Golgi Network. DAPI: 4′,6-diamidino-2-phenylindole
Purified 4 Mouse Anti Nf Atc3 Monoclonal Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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purified 4 mouse anti-nf-atc3 monoclonal antibody - by Bioz Stars, 2026-10
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biotinylated anti-active caspase-3 rabbit monoclonal antibody  (Becton Dickinson)
90
Becton Dickinson biotinylated anti-active caspase-3 rabbit monoclonal antibody
Figure 1 Vector construction, validation and assessment of <t>EphA2</t> expression. (A) Schematic of the C170 and C172 EphA2 expression viruses. (B) Southern blot confirming the anticipated NcoI fragments in our new recombinants (1.2 and 3.0 kb, C170; 2.3 and 3.0 kb, C172), (C) Immunofluorescent imaging shows different cellular distributions of the full length and secreted forms of the EphA2. (D) Western blots of CT2A infected cells and supernatants show that C170-expressed EphA2 remains cell associated, whereas C172 expressing the extracellular form of EphA2 secretes the protein into the supernatant. TAA: tumor- associated antigen. UTR: untranslated region. TGN: Trans Golgi Network. DAPI: 4′,6-diamidino-2-phenylindole
Biotinylated Anti Active Caspase 3 Rabbit Monoclonal Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+nf+%CE%BAb/mouse+monoclonal+anti+nf+kb+p65++ps536++j144+460+ruo/pmc02636855-71-10-37
Average 90 stars, based on 1 article reviews
biotinylated anti-active caspase-3 rabbit monoclonal antibody - by Bioz Stars, 2026-10
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Image Search Results


Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.

Journal: bioRxiv

Article Title: Selective Immune Silencing by Targeted TGF-β Agonists

doi: 10.64898/2026.01.19.700410

Figure Lengend Snippet: Design and characterization of mouse T cell-targeted TGF- β agonists. a. Schematic of the plasmid construct encoding the MSA–TGM1-targeting-arm fusion protein. b. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. c. SDS–PAGE analysis showing the molecular weight and purity of the indicated recombinant proteins. d. Representative flow cytometry plots showing mouse CD4 and CD8α expression in mouse CD4- or CD8α-transduced HEK293 cells. e. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. f. Dose-dependent curves showing GITR, IRF4, and T-bet expression levels in in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. g. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD4 T cells treated with the indicated proteins on day 3. h. Dose-dependent curves showing percentages of IFN-γ– and TNF-α–producing cells among in vitro –cultured CD8 T cells treated with the indicated proteins on day 3. i. Dose-dependent curves showing Foxp3 cell percentages in in vitro -cultured CD4 T cells, and Granzyme B–producing cell percentages in in vitro -cultured CD8 T cells treated with the indicated proteins on day 3. Data are presented as mean ± s.e.m.

Article Snippet: The following antibodies were purchased from Miltenyi Biotec: human IgA (130-113-480) and IRF4 (130-100-909).

Techniques: Plasmid Preparation, Construct, SDS Page, Molecular Weight, Recombinant, Flow Cytometry, Expressing, In Vitro, Cell Culture

a. Frequencies of immune cell subsets. b. SCENIC prediction of IRF4 and Blimp1 transcription factor activity. c. Immunoglobulin gene expression. d. UMAP plots showing cell cycle gene signature scores. e. Quantification of cell cycle gene signature scores. f. Scaled signature scores for TGF-β–inducible genes, senescence genes, and apoptosis-related genes, aggregated by sample.

Journal: bioRxiv

Article Title: Selective Immune Silencing by Targeted TGF-β Agonists

doi: 10.64898/2026.01.19.700410

Figure Lengend Snippet: a. Frequencies of immune cell subsets. b. SCENIC prediction of IRF4 and Blimp1 transcription factor activity. c. Immunoglobulin gene expression. d. UMAP plots showing cell cycle gene signature scores. e. Quantification of cell cycle gene signature scores. f. Scaled signature scores for TGF-β–inducible genes, senescence genes, and apoptosis-related genes, aggregated by sample.

Article Snippet: The following antibodies were purchased from Miltenyi Biotec: human IgA (130-113-480) and IRF4 (130-100-909).

Techniques: Activity Assay, Gene Expression

Figure 1 Vector construction, validation and assessment of EphA2 expression. (A) Schematic of the C170 and C172 EphA2 expression viruses. (B) Southern blot confirming the anticipated NcoI fragments in our new recombinants (1.2 and 3.0 kb, C170; 2.3 and 3.0 kb, C172), (C) Immunofluorescent imaging shows different cellular distributions of the full length and secreted forms of the EphA2. (D) Western blots of CT2A infected cells and supernatants show that C170-expressed EphA2 remains cell associated, whereas C172 expressing the extracellular form of EphA2 secretes the protein into the supernatant. TAA: tumor- associated antigen. UTR: untranslated region. TGN: Trans Golgi Network. DAPI: 4′,6-diamidino-2-phenylindole

Journal: Journal for immunotherapy of cancer

Article Title: Eliciting an immune-mediated antitumor response through oncolytic herpes simplex virus-based shared antigen expression in tumors resistant to viroimmunotherapy.

doi: 10.1136/jitc-2021-002939

Figure Lengend Snippet: Figure 1 Vector construction, validation and assessment of EphA2 expression. (A) Schematic of the C170 and C172 EphA2 expression viruses. (B) Southern blot confirming the anticipated NcoI fragments in our new recombinants (1.2 and 3.0 kb, C170; 2.3 and 3.0 kb, C172), (C) Immunofluorescent imaging shows different cellular distributions of the full length and secreted forms of the EphA2. (D) Western blots of CT2A infected cells and supernatants show that C170-expressed EphA2 remains cell associated, whereas C172 expressing the extracellular form of EphA2 secretes the protein into the supernatant. TAA: tumor- associated antigen. UTR: untranslated region. TGN: Trans Golgi Network. DAPI: 4′,6-diamidino-2-phenylindole

Article Snippet: Cells were labeled with the following antibody staining panels for analysis of the adaptive immune cells: CD11b- Violet 421 (M1/70), CD4- BV785 (GK1.5), CD25- PE (7D4/CD25), CD8aBV510 (53–6.7), CD3ε-BV 711 (145–2 C11), CD44- APC (IM7), CD45- BV605 (30- F11), NKp46–PE- Cy7 (29A1.4) and B220- AF488 (RA3- 6B2), and H2Kb/H2Db (28- 8- 6) from Bio- Legend (San Diego, California, USA) and mouse EphA2 (REA579) from Miltenyi Biotec (Auburn, California, USA).

Techniques: Plasmid Preparation, Biomarker Discovery, Expressing, Southern Blot, Imaging, Western Blot, Infection

Figure 2 Viral replication and cytopathic effect in C57BL/6 murine CT2A MG panels show viral replication kinetics, cytopathic effect and EphA2 surface expression in infected CT2A C57BL/6-based MG cells. MG, malignant glioma.

Journal: Journal for immunotherapy of cancer

Article Title: Eliciting an immune-mediated antitumor response through oncolytic herpes simplex virus-based shared antigen expression in tumors resistant to viroimmunotherapy.

doi: 10.1136/jitc-2021-002939

Figure Lengend Snippet: Figure 2 Viral replication and cytopathic effect in C57BL/6 murine CT2A MG panels show viral replication kinetics, cytopathic effect and EphA2 surface expression in infected CT2A C57BL/6-based MG cells. MG, malignant glioma.

Article Snippet: Cells were labeled with the following antibody staining panels for analysis of the adaptive immune cells: CD11b- Violet 421 (M1/70), CD4- BV785 (GK1.5), CD25- PE (7D4/CD25), CD8aBV510 (53–6.7), CD3ε-BV 711 (145–2 C11), CD44- APC (IM7), CD45- BV605 (30- F11), NKp46–PE- Cy7 (29A1.4) and B220- AF488 (RA3- 6B2), and H2Kb/H2Db (28- 8- 6) from Bio- Legend (San Diego, California, USA) and mouse EphA2 (REA579) from Miltenyi Biotec (Auburn, California, USA).

Techniques: Expressing, Infection

Figure 4 CT2A tumor infiltrate immunophenotypic analysis. (A). Representative summary of TILs and population changes 6 days post saline, C134, and C170 (C134 +Epha2) treatment. Numbers below pie chart represent TILs/ml brain. (B–D). T-cell tumor infiltrate and subset analysis from saline (green), C134 (salmon), and C170 (blue)-treated mice. (E) Representative gating summary of CD8 subsets and CD62L/CD44 staining in treated mice. CD8 subset analysis of (F) CD8, CD25+, (G) CD8+, CD44+, CD62L effector-like population changes, and (H) CD8+, CD44+, CD62L+central memory-like population changes in C170- treated mice. Data were analyzed by one-way analysis of variance; p values were adjusted by Holm’s procedure. TIL: tumor- infiltrating leukocyte, MDSC: myeloid derived suppressor cells.

Journal: Journal for immunotherapy of cancer

Article Title: Eliciting an immune-mediated antitumor response through oncolytic herpes simplex virus-based shared antigen expression in tumors resistant to viroimmunotherapy.

doi: 10.1136/jitc-2021-002939

Figure Lengend Snippet: Figure 4 CT2A tumor infiltrate immunophenotypic analysis. (A). Representative summary of TILs and population changes 6 days post saline, C134, and C170 (C134 +Epha2) treatment. Numbers below pie chart represent TILs/ml brain. (B–D). T-cell tumor infiltrate and subset analysis from saline (green), C134 (salmon), and C170 (blue)-treated mice. (E) Representative gating summary of CD8 subsets and CD62L/CD44 staining in treated mice. CD8 subset analysis of (F) CD8, CD25+, (G) CD8+, CD44+, CD62L effector-like population changes, and (H) CD8+, CD44+, CD62L+central memory-like population changes in C170- treated mice. Data were analyzed by one-way analysis of variance; p values were adjusted by Holm’s procedure. TIL: tumor- infiltrating leukocyte, MDSC: myeloid derived suppressor cells.

Article Snippet: Cells were labeled with the following antibody staining panels for analysis of the adaptive immune cells: CD11b- Violet 421 (M1/70), CD4- BV785 (GK1.5), CD25- PE (7D4/CD25), CD8aBV510 (53–6.7), CD3ε-BV 711 (145–2 C11), CD44- APC (IM7), CD45- BV605 (30- F11), NKp46–PE- Cy7 (29A1.4) and B220- AF488 (RA3- 6B2), and H2Kb/H2Db (28- 8- 6) from Bio- Legend (San Diego, California, USA) and mouse EphA2 (REA579) from Miltenyi Biotec (Auburn, California, USA).

Techniques: Saline, Staining, Derivative Assay

Figure 7 Summary of 67C-4-oHSV antitumor response studies. C170 was evaluated in vitro and showed equivalent (A) replication and (B) cytopathic activity as the parent virus C134. (C) 67 C-4 expresses EphA2 on the cell surface in mock and C170 infected cells. (D) Schematic summary of experimental approach. (E&F) Independent studies show C170 virotherapy treatment of established 67 C-4 flank tumors shows that C170 significantly suppresses tumor growth when compared with saline or C134-treated cohorts. Data were analyzed by using analysis of variance with repeated measures. P values were adjusted by Holm’s procedure.

Journal: Journal for immunotherapy of cancer

Article Title: Eliciting an immune-mediated antitumor response through oncolytic herpes simplex virus-based shared antigen expression in tumors resistant to viroimmunotherapy.

doi: 10.1136/jitc-2021-002939

Figure Lengend Snippet: Figure 7 Summary of 67C-4-oHSV antitumor response studies. C170 was evaluated in vitro and showed equivalent (A) replication and (B) cytopathic activity as the parent virus C134. (C) 67 C-4 expresses EphA2 on the cell surface in mock and C170 infected cells. (D) Schematic summary of experimental approach. (E&F) Independent studies show C170 virotherapy treatment of established 67 C-4 flank tumors shows that C170 significantly suppresses tumor growth when compared with saline or C134-treated cohorts. Data were analyzed by using analysis of variance with repeated measures. P values were adjusted by Holm’s procedure.

Article Snippet: Cells were labeled with the following antibody staining panels for analysis of the adaptive immune cells: CD11b- Violet 421 (M1/70), CD4- BV785 (GK1.5), CD25- PE (7D4/CD25), CD8aBV510 (53–6.7), CD3ε-BV 711 (145–2 C11), CD44- APC (IM7), CD45- BV605 (30- F11), NKp46–PE- Cy7 (29A1.4) and B220- AF488 (RA3- 6B2), and H2Kb/H2Db (28- 8- 6) from Bio- Legend (San Diego, California, USA) and mouse EphA2 (REA579) from Miltenyi Biotec (Auburn, California, USA).

Techniques: In Vitro, Activity Assay, Virus, Infection, Saline

Figure 8 T-cell function studies from saline and oHSV treated mice shows that C170 treatment induces an antigen-specific T- cell response in the periphery of long-term survivors. Splenocytes from saline (blue column) or oHSV-treated mice (red column, C134; green column, C170) were analyzed. (A) At the start, there was no difference in the populations that used peptide pulsing; however, after pulsing with 10 µm EphA2 or 10µM OVA peptide (negative control), C170-treated mice significantly increase their activated (B) CD25(+), (C) GZMB(+), and (D) CD25+, GZMB + dual staining CD8 + populations indicative of an EphA2-specific population response. (E) Representative flow plot showingCD8(+) GZMB(+) population and gating, and the (F) GZMB CD25 dual-positive populations. oHSV, oncolytic herpes simplex virus; OVA, ovalbumin.

Journal: Journal for immunotherapy of cancer

Article Title: Eliciting an immune-mediated antitumor response through oncolytic herpes simplex virus-based shared antigen expression in tumors resistant to viroimmunotherapy.

doi: 10.1136/jitc-2021-002939

Figure Lengend Snippet: Figure 8 T-cell function studies from saline and oHSV treated mice shows that C170 treatment induces an antigen-specific T- cell response in the periphery of long-term survivors. Splenocytes from saline (blue column) or oHSV-treated mice (red column, C134; green column, C170) were analyzed. (A) At the start, there was no difference in the populations that used peptide pulsing; however, after pulsing with 10 µm EphA2 or 10µM OVA peptide (negative control), C170-treated mice significantly increase their activated (B) CD25(+), (C) GZMB(+), and (D) CD25+, GZMB + dual staining CD8 + populations indicative of an EphA2-specific population response. (E) Representative flow plot showingCD8(+) GZMB(+) population and gating, and the (F) GZMB CD25 dual-positive populations. oHSV, oncolytic herpes simplex virus; OVA, ovalbumin.

Article Snippet: Cells were labeled with the following antibody staining panels for analysis of the adaptive immune cells: CD11b- Violet 421 (M1/70), CD4- BV785 (GK1.5), CD25- PE (7D4/CD25), CD8aBV510 (53–6.7), CD3ε-BV 711 (145–2 C11), CD44- APC (IM7), CD45- BV605 (30- F11), NKp46–PE- Cy7 (29A1.4) and B220- AF488 (RA3- 6B2), and H2Kb/H2Db (28- 8- 6) from Bio- Legend (San Diego, California, USA) and mouse EphA2 (REA579) from Miltenyi Biotec (Auburn, California, USA).

Techniques: Cell Function Assay, Saline, Negative Control, Staining, Virus