mouse anti nf Search Results


irf4 rea201  (Miltenyi Biotec)
94
Miltenyi Biotec irf4 rea201
Figure 4. <t>IRF4</t> and NFAT Form a Positive Feedback Circuit during T Cell Exhaustion (A) Expression of Nfatc1 isoforms as identified by RNA sequencing. Graph shows expression relative to naive T cells of the short isoform (NFATc1aA, represented by exon 1–3 junctions, skipping exon 2) and the long isoform (represented by exon 2–3 junctions) in P14 T cells isolated from acutely (WE) and chronically (Docile) LCMV-infected mice at day 30 post infection (p.i.). (B) Western blot showing NFATc1, NFATc2, IRF4, BATF, and Actin (loading control) expression in CD8+ T cells flow cytometry sorted as CD62L from acutely and as CD62LPD-1+ from chronically LCMV-infected mice at day 30 p.i. The arrow marks the short isoform of NFATc1. (C) Binding of NFATc1 and NFATc2 to the Irf4 and Pdcd1 (encoding PD-1) promoters demonstrated by chromatin immunoprecipitation using specific antibodies or control IgG and RT-qPCR in total CD44+CD8+ T cells isolated from spleens of acutely and chronically LCMV-infected mice at day 8 p.i. Enrichment is expressed as percentage of total chromatin input and compares isotype control and NFATc1- and NFATc2-specific antibodies. (D) Western blot showing NFATc1, NFATc2, IRF4, BATF, and p50 NF-kB (loading control) expression in polyclonal CD8+ T cells deficient (KO) in NFATc1 (Nfatc1fl/flCd4Cre), NFATc2 (Nfatc2/), or both (DKO) after in vitro activation with anti-CD3 and anti-CD28 for 48 hr. (E) Western blot showing NFATc1, IRF4, BATF, and Actin (loading control) in polyclonal CD8+ T cells activated with anti-CD3, anti-CD28, and IL-2 with or without cyclosporine A (CsA) for 24 hr. Data in (A)–(C) are representative of 2 independent experiments and data in (D) and (E) are representative of 3 independent experiments. Error bars denote mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t test (*p < 0.05, **p < 0.01, ***p < 0.001).
Irf4 Rea201, 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/pm29246443-247-57-60?v=Miltenyi+Biotec
Average 94 stars, based on 1 article reviews
irf4 rea201 - by Bioz Stars, 2026-07
94/100 stars
  Buy from Supplier
akt 281046  (R&D Systems)
93
R&D Systems akt 281046
Figure 4. <t>IRF4</t> and NFAT Form a Positive Feedback Circuit during T Cell Exhaustion (A) Expression of Nfatc1 isoforms as identified by RNA sequencing. Graph shows expression relative to naive T cells of the short isoform (NFATc1aA, represented by exon 1–3 junctions, skipping exon 2) and the long isoform (represented by exon 2–3 junctions) in P14 T cells isolated from acutely (WE) and chronically (Docile) LCMV-infected mice at day 30 post infection (p.i.). (B) Western blot showing NFATc1, NFATc2, IRF4, BATF, and Actin (loading control) expression in CD8+ T cells flow cytometry sorted as CD62L from acutely and as CD62LPD-1+ from chronically LCMV-infected mice at day 30 p.i. The arrow marks the short isoform of NFATc1. (C) Binding of NFATc1 and NFATc2 to the Irf4 and Pdcd1 (encoding PD-1) promoters demonstrated by chromatin immunoprecipitation using specific antibodies or control IgG and RT-qPCR in total CD44+CD8+ T cells isolated from spleens of acutely and chronically LCMV-infected mice at day 8 p.i. Enrichment is expressed as percentage of total chromatin input and compares isotype control and NFATc1- and NFATc2-specific antibodies. (D) Western blot showing NFATc1, NFATc2, IRF4, BATF, and p50 NF-kB (loading control) expression in polyclonal CD8+ T cells deficient (KO) in NFATc1 (Nfatc1fl/flCd4Cre), NFATc2 (Nfatc2/), or both (DKO) after in vitro activation with anti-CD3 and anti-CD28 for 48 hr. (E) Western blot showing NFATc1, IRF4, BATF, and Actin (loading control) in polyclonal CD8+ T cells activated with anti-CD3, anti-CD28, and IL-2 with or without cyclosporine A (CsA) for 24 hr. Data in (A)–(C) are representative of 2 independent experiments and data in (D) and (E) are representative of 3 independent experiments. Error bars denote mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t test (*p < 0.05, **p < 0.01, ***p < 0.001).
Akt 281046, 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/pm37457144-41-14-34?v=R%26D+Systems
Average 93 stars, based on 1 article reviews
akt 281046 - by Bioz Stars, 2026-07
93/100 stars
  Buy from Supplier
anti rela  (R&D Systems)
94
R&D Systems anti rela
Figure 4. <t>IRF4</t> and NFAT Form a Positive Feedback Circuit during T Cell Exhaustion (A) Expression of Nfatc1 isoforms as identified by RNA sequencing. Graph shows expression relative to naive T cells of the short isoform (NFATc1aA, represented by exon 1–3 junctions, skipping exon 2) and the long isoform (represented by exon 2–3 junctions) in P14 T cells isolated from acutely (WE) and chronically (Docile) LCMV-infected mice at day 30 post infection (p.i.). (B) Western blot showing NFATc1, NFATc2, IRF4, BATF, and Actin (loading control) expression in CD8+ T cells flow cytometry sorted as CD62L from acutely and as CD62LPD-1+ from chronically LCMV-infected mice at day 30 p.i. The arrow marks the short isoform of NFATc1. (C) Binding of NFATc1 and NFATc2 to the Irf4 and Pdcd1 (encoding PD-1) promoters demonstrated by chromatin immunoprecipitation using specific antibodies or control IgG and RT-qPCR in total CD44+CD8+ T cells isolated from spleens of acutely and chronically LCMV-infected mice at day 8 p.i. Enrichment is expressed as percentage of total chromatin input and compares isotype control and NFATc1- and NFATc2-specific antibodies. (D) Western blot showing NFATc1, NFATc2, IRF4, BATF, and p50 NF-kB (loading control) expression in polyclonal CD8+ T cells deficient (KO) in NFATc1 (Nfatc1fl/flCd4Cre), NFATc2 (Nfatc2/), or both (DKO) after in vitro activation with anti-CD3 and anti-CD28 for 48 hr. (E) Western blot showing NFATc1, IRF4, BATF, and Actin (loading control) in polyclonal CD8+ T cells activated with anti-CD3, anti-CD28, and IL-2 with or without cyclosporine A (CsA) for 24 hr. Data in (A)–(C) are representative of 2 independent experiments and data in (D) and (E) are representative of 3 independent experiments. Error bars denote mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t test (*p < 0.05, **p < 0.01, ***p < 0.001).
Anti Rela, 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/pmc05833376-235-7-12?v=R%26D+Systems
Average 94 stars, based on 1 article reviews
anti rela - by Bioz Stars, 2026-07
94/100 stars
  Buy from Supplier
b v co  (Miltenyi Biotec)
94
Miltenyi Biotec b v co
Figure 4. <t>IRF4</t> and NFAT Form a Positive Feedback Circuit during T Cell Exhaustion (A) Expression of Nfatc1 isoforms as identified by RNA sequencing. Graph shows expression relative to naive T cells of the short isoform (NFATc1aA, represented by exon 1–3 junctions, skipping exon 2) and the long isoform (represented by exon 2–3 junctions) in P14 T cells isolated from acutely (WE) and chronically (Docile) LCMV-infected mice at day 30 post infection (p.i.). (B) Western blot showing NFATc1, NFATc2, IRF4, BATF, and Actin (loading control) expression in CD8+ T cells flow cytometry sorted as CD62L from acutely and as CD62LPD-1+ from chronically LCMV-infected mice at day 30 p.i. The arrow marks the short isoform of NFATc1. (C) Binding of NFATc1 and NFATc2 to the Irf4 and Pdcd1 (encoding PD-1) promoters demonstrated by chromatin immunoprecipitation using specific antibodies or control IgG and RT-qPCR in total CD44+CD8+ T cells isolated from spleens of acutely and chronically LCMV-infected mice at day 8 p.i. Enrichment is expressed as percentage of total chromatin input and compares isotype control and NFATc1- and NFATc2-specific antibodies. (D) Western blot showing NFATc1, NFATc2, IRF4, BATF, and p50 NF-kB (loading control) expression in polyclonal CD8+ T cells deficient (KO) in NFATc1 (Nfatc1fl/flCd4Cre), NFATc2 (Nfatc2/), or both (DKO) after in vitro activation with anti-CD3 and anti-CD28 for 48 hr. (E) Western blot showing NFATc1, IRF4, BATF, and Actin (loading control) in polyclonal CD8+ T cells activated with anti-CD3, anti-CD28, and IL-2 with or without cyclosporine A (CsA) for 24 hr. Data in (A)–(C) are representative of 2 independent experiments and data in (D) and (E) are representative of 3 independent experiments. Error bars denote mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t test (*p < 0.05, **p < 0.01, ***p < 0.001).
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/pmc12796729-24-8-6?v=Miltenyi+Biotec
Average 94 stars, based on 1 article reviews
b v co - by Bioz Stars, 2026-07
94/100 stars
  Buy from Supplier
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/pm34599026-83-60-64?v=Miltenyi+Biotec
Average 94 stars, based on 1 article reviews
mouse epha2 rea579 - by Bioz Stars, 2026-07
94/100 stars
  Buy from Supplier
oct3 4 apc  (Miltenyi Biotec)
94
Miltenyi Biotec oct3 4 apc
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
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/pm38771392-363-45-47?v=Miltenyi+Biotec
Average 94 stars, based on 1 article reviews
oct3 4 apc - by Bioz Stars, 2026-07
94/100 stars
  Buy from Supplier
7074 anti total p65 r d systems  (R&D Systems)
92
R&D Systems 7074 anti total p65 r d systems
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
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/pmc10986120__13046_2024_3019_MOESM2_ESM-35-2-5?v=R%26D+Systems
Average 92 stars, based on 1 article reviews
7074 anti total p65 r d systems - by Bioz Stars, 2026-07
92/100 stars
  Buy from Supplier
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/pmc06605374-328-17-23?v=Rockland+Immunochemicals
Average 85 stars, based on 1 article reviews
axons - by Bioz Stars, 2026-07
85/100 stars
  Buy from Supplier
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/pmc03758556-199-17-24?v=Bio-Rad
Average 86 stars, based on 1 article reviews
mouse anti nf antibody - by Bioz Stars, 2026-07
86/100 stars
  Buy from Supplier
anti-nf-α1/cpe monoclonal antibody  (Becton Dickinson)
90
Becton Dickinson anti-nf-α1/cpe 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
Anti Nf α1/Cpe 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/pm34966948-65-21-24?v=Becton+Dickinson
Average 90 stars, based on 1 article reviews
anti-nf-α1/cpe monoclonal antibody - by Bioz Stars, 2026-07
90/100 stars
  Buy from Supplier
alexa 647-conjugated phospho-nf-kb p65 (rela) antibody  (Becton Dickinson)
90
Becton Dickinson alexa 647-conjugated phospho-nf-kb p65 (rela) 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
Alexa 647 Conjugated Phospho Nf Kb P65 (Rela) 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/10__1158_slash_1535___7163__mct___10___0829-45-21-26?v=Becton+Dickinson
Average 90 stars, based on 1 article reviews
alexa 647-conjugated phospho-nf-kb p65 (rela) antibody - by Bioz Stars, 2026-07
90/100 stars
  Buy from Supplier
mouse anti-200 kda-nf  (ICN Biomedicals)
90
ICN Biomedicals mouse anti-200 kda-nf
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 200 Kda Nf, supplied by ICN Biomedicals, 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/10__1523_slash_jneurosci__5080___09__2010-118-35-39?v=ICN+Biomedicals
Average 90 stars, based on 1 article reviews
mouse anti-200 kda-nf - by Bioz Stars, 2026-07
90/100 stars
  Buy from Supplier

Image Search Results


Figure 4. IRF4 and NFAT Form a Positive Feedback Circuit during T Cell Exhaustion (A) Expression of Nfatc1 isoforms as identified by RNA sequencing. Graph shows expression relative to naive T cells of the short isoform (NFATc1aA, represented by exon 1–3 junctions, skipping exon 2) and the long isoform (represented by exon 2–3 junctions) in P14 T cells isolated from acutely (WE) and chronically (Docile) LCMV-infected mice at day 30 post infection (p.i.). (B) Western blot showing NFATc1, NFATc2, IRF4, BATF, and Actin (loading control) expression in CD8+ T cells flow cytometry sorted as CD62L from acutely and as CD62LPD-1+ from chronically LCMV-infected mice at day 30 p.i. The arrow marks the short isoform of NFATc1. (C) Binding of NFATc1 and NFATc2 to the Irf4 and Pdcd1 (encoding PD-1) promoters demonstrated by chromatin immunoprecipitation using specific antibodies or control IgG and RT-qPCR in total CD44+CD8+ T cells isolated from spleens of acutely and chronically LCMV-infected mice at day 8 p.i. Enrichment is expressed as percentage of total chromatin input and compares isotype control and NFATc1- and NFATc2-specific antibodies. (D) Western blot showing NFATc1, NFATc2, IRF4, BATF, and p50 NF-kB (loading control) expression in polyclonal CD8+ T cells deficient (KO) in NFATc1 (Nfatc1fl/flCd4Cre), NFATc2 (Nfatc2/), or both (DKO) after in vitro activation with anti-CD3 and anti-CD28 for 48 hr. (E) Western blot showing NFATc1, IRF4, BATF, and Actin (loading control) in polyclonal CD8+ T cells activated with anti-CD3, anti-CD28, and IL-2 with or without cyclosporine A (CsA) for 24 hr. Data in (A)–(C) are representative of 2 independent experiments and data in (D) and (E) are representative of 3 independent experiments. Error bars denote mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t test (*p < 0.05, **p < 0.01, ***p < 0.001).

Journal: Immunity

Article Title: Transcription Factor IRF4 Promotes CD8 + T Cell Exhaustion and Limits the Development of Memory-like T Cells during Chronic Infection.

doi: 10.1016/j.immuni.2017.11.021

Figure Lengend Snippet: Figure 4. IRF4 and NFAT Form a Positive Feedback Circuit during T Cell Exhaustion (A) Expression of Nfatc1 isoforms as identified by RNA sequencing. Graph shows expression relative to naive T cells of the short isoform (NFATc1aA, represented by exon 1–3 junctions, skipping exon 2) and the long isoform (represented by exon 2–3 junctions) in P14 T cells isolated from acutely (WE) and chronically (Docile) LCMV-infected mice at day 30 post infection (p.i.). (B) Western blot showing NFATc1, NFATc2, IRF4, BATF, and Actin (loading control) expression in CD8+ T cells flow cytometry sorted as CD62L from acutely and as CD62LPD-1+ from chronically LCMV-infected mice at day 30 p.i. The arrow marks the short isoform of NFATc1. (C) Binding of NFATc1 and NFATc2 to the Irf4 and Pdcd1 (encoding PD-1) promoters demonstrated by chromatin immunoprecipitation using specific antibodies or control IgG and RT-qPCR in total CD44+CD8+ T cells isolated from spleens of acutely and chronically LCMV-infected mice at day 8 p.i. Enrichment is expressed as percentage of total chromatin input and compares isotype control and NFATc1- and NFATc2-specific antibodies. (D) Western blot showing NFATc1, NFATc2, IRF4, BATF, and p50 NF-kB (loading control) expression in polyclonal CD8+ T cells deficient (KO) in NFATc1 (Nfatc1fl/flCd4Cre), NFATc2 (Nfatc2/), or both (DKO) after in vitro activation with anti-CD3 and anti-CD28 for 48 hr. (E) Western blot showing NFATc1, IRF4, BATF, and Actin (loading control) in polyclonal CD8+ T cells activated with anti-CD3, anti-CD28, and IL-2 with or without cyclosporine A (CsA) for 24 hr. Data in (A)–(C) are representative of 2 independent experiments and data in (D) and (E) are representative of 3 independent experiments. Error bars denote mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t test (*p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: Fluorochrome-conjugated antibodies directed against the following antigens were used for analysis by flow cytometry: CD8a (53-6.7), CD62L (MEL-14), CD71 (R17217), CD98 (RL388), Ly5.1 (A20), 2B4 (eBio244F4), PD-1 (J43), Lag3 (T47-530), TIGIT (GIGD7), CTLA4 (14D3), TIM-3 (RMT3-23), CD127 (A7R34), CXCR5 (SPRCL5), IL-2 (JES6-5H4), IFNg (XMG1.2) (from Thermo Fisher Scientific), CD44 (IM7), Ly5.2 (104), TNF (MP5-XT22) (from BD Biosciences), IRF4 (REA201) (from Miltenyi), TCF1 (C63D9), BATF (D7C5) (from Cell Signaling Technology), IRF4 (M17) (from Santa Cruz Biotechnology), and anti-goat IgG coupled to fluorescein isothiocyanate (Jackson ImmunoResearch Laboratories) and anti-rabbit IgG coupled to Alexa Fluor 647 (Thermo Fisher Scientific). e3 Immunity 47, 1–13.e1–e5, December 19, 2017 Propidium iodide, SytoxBlue or fixable viability dye eFluor506 (all Thermo Fisher Scientific) were used to exclude dead cells.

Techniques: Expressing, RNA Sequencing, Isolation, Infection, Western Blot, Control, Cytometry, Binding Assay, Chromatin Immunoprecipitation, Quantitative RT-PCR, In Vitro, Activation Assay, Two Tailed Test

Figure 5. IRF4 and BATF Cooperate with NFAT to Establish T Cell Exhaustion (A) Venn diagram showing overlap between genes bound by IRF4, BATF, and NFAT as identified by chromatin immunoprecipitation (ChIP) sequencing of effector CD8+ T cells (Kurachi et al., 2014; Man et al., 2013; Martinez et al., 2015). (legend continued on next page) Immunity 47, 1–13, December 19, 2017 7

Journal: Immunity

Article Title: Transcription Factor IRF4 Promotes CD8 + T Cell Exhaustion and Limits the Development of Memory-like T Cells during Chronic Infection.

doi: 10.1016/j.immuni.2017.11.021

Figure Lengend Snippet: Figure 5. IRF4 and BATF Cooperate with NFAT to Establish T Cell Exhaustion (A) Venn diagram showing overlap between genes bound by IRF4, BATF, and NFAT as identified by chromatin immunoprecipitation (ChIP) sequencing of effector CD8+ T cells (Kurachi et al., 2014; Man et al., 2013; Martinez et al., 2015). (legend continued on next page) Immunity 47, 1–13, December 19, 2017 7

Article Snippet: Fluorochrome-conjugated antibodies directed against the following antigens were used for analysis by flow cytometry: CD8a (53-6.7), CD62L (MEL-14), CD71 (R17217), CD98 (RL388), Ly5.1 (A20), 2B4 (eBio244F4), PD-1 (J43), Lag3 (T47-530), TIGIT (GIGD7), CTLA4 (14D3), TIM-3 (RMT3-23), CD127 (A7R34), CXCR5 (SPRCL5), IL-2 (JES6-5H4), IFNg (XMG1.2) (from Thermo Fisher Scientific), CD44 (IM7), Ly5.2 (104), TNF (MP5-XT22) (from BD Biosciences), IRF4 (REA201) (from Miltenyi), TCF1 (C63D9), BATF (D7C5) (from Cell Signaling Technology), IRF4 (M17) (from Santa Cruz Biotechnology), and anti-goat IgG coupled to fluorescein isothiocyanate (Jackson ImmunoResearch Laboratories) and anti-rabbit IgG coupled to Alexa Fluor 647 (Thermo Fisher Scientific). e3 Immunity 47, 1–13.e1–e5, December 19, 2017 Propidium iodide, SytoxBlue or fixable viability dye eFluor506 (all Thermo Fisher Scientific) were used to exclude dead cells.

Techniques: Chromatin Immunoprecipitation, ChIP-sequencing

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