mouse tumor cells Search Results


mouse leydig cell tumor  (ATCC)
94
ATCC mouse leydig cell tumor
Mouse Leydig Cell Tumor, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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tumor isolation kit  (Miltenyi Biotec)
95
Miltenyi Biotec tumor isolation kit
Tumor Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse monoclonal kif5b antibody  (Rockland Immunochemicals)
90
Rockland Immunochemicals mouse monoclonal kif5b antibody
a Relative quantification of Kif5a, <t>Kif5b</t> and Kif5c transcripts by real-time PCR in CD8α + and CD11b + DCs and BMDCs from WT or cKO Kif5b mice. Transcript levels for each sample are expressed relative to Kif5b. Graph shows mean ± S.E.M. ( n = 3). b Contour plots of DCs from the spleen of either WT or cKO Kif5b mice pregated on CD19 − , Gr − , F4/80 − CD11c + , and IA/IE + . The data are representative of three independent experiments. c Absolute numbers of CD11b + DCs and CD8α + DCs in the spleen of WT (blue circles) and cKO Kif5b (red squares) mice. The data are representative of five independent experiments. d – g The efficiency of cross-presentation of different concentration of sOVA and the presentation of OVA peptide SIINFEKL in vitro by CD8α + DCs ( d , g ), CD11b + DCs ( e , g ) and BMDCs ( f , g ) from WT (blue histogram or line) or cKO Kif5b (red histogram or line) mice was measured as IL-2 secretion by OT-I T cells after 16 h of co-culture. Graph shows mean ± S.E.M. ( n = 4). Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. * P < 0.05; ** P < 0.005; *** P < 0.0001.
Mouse Monoclonal Kif5b Antibody, supplied by Rockland Immunochemicals, 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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recombinant mouse tnfα  (Cell Applications Inc)
93
Cell Applications Inc recombinant mouse tnfα
Fig. 4 Conserved NF-κB binding is shared between cell types near common immune response genes. a Representative genomic regions showing the pan-cell (four-cell-type-shared: between human aortic ECs [HAECs], human umbilical vein ECs [HUVECs], lymphoblastoid cell lines [LCLs], and adipocytes), EC-specific (shared only between HAECs and HUVECs), LCL-specific, and adipocyte-specific RELA peaks. b Stacked bar charts showing the fractions of four-, three-, and two-cell-type-shared and HAEC-specific RELA peaks (one-cell type) that are three-species-conserved, two-species- conserved, or human-specific. c Table showing the top-scoring de novo motifs (MEME-ChIP e values, Supplementary Dataset 4), fold enrichments of RELA binding modes (p value: *<1.0 × 10−6, Chi-squared test with Yates’s correction for continuity of independence and Bonferroni correction for multiple testing), fold enrichments near <t>TNFα</t> target genes (HAEC RNA-seq, ±10 kb of transcriptional start sites, p value: * <1.0 × 10−5, two-sided Fisher’s exact test with Bonferroni correction), and the top GO terms (GREAT FDR q values, Biological Process Ontology, Supplementary Dataset 3) for the pan-cell (four-cell- type-shared), EC-specific, and HAEC-specific RELA peaks. ↑= upregulated, ↓= downregulated, NC no change (i.e., constitutively expressed). d Profile plots comparing ERG binding signal at pan-cell, EC-specific, and HAEC-specific RELA peaks in HAECs. ERG ChIP-seq signal was generated using published raw data from ref. 15. The plot lines indicate mean RPM ± SEM. Arrows and percentages indicate change in signal after TNFα stimulation. The p values were calculated using two-sided Welch Two Sample t-test (p value: *<0.05). e, Model of NF-κB-ERG dynamics and cofactor squelching showing redistribution of cofactors from EC-specific Mode O regions to pan-cell Mode P regions. Source data are provided in the Source Data file for Fig. 4c, d.
Recombinant Mouse Tnfα, supplied by Cell Applications Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse brain tumor cell line kt 98  (ATCC)
90
ATCC mouse brain tumor cell line kt 98
Fig. 4 Conserved NF-κB binding is shared between cell types near common immune response genes. a Representative genomic regions showing the pan-cell (four-cell-type-shared: between human aortic ECs [HAECs], human umbilical vein ECs [HUVECs], lymphoblastoid cell lines [LCLs], and adipocytes), EC-specific (shared only between HAECs and HUVECs), LCL-specific, and adipocyte-specific RELA peaks. b Stacked bar charts showing the fractions of four-, three-, and two-cell-type-shared and HAEC-specific RELA peaks (one-cell type) that are three-species-conserved, two-species- conserved, or human-specific. c Table showing the top-scoring de novo motifs (MEME-ChIP e values, Supplementary Dataset 4), fold enrichments of RELA binding modes (p value: *<1.0 × 10−6, Chi-squared test with Yates’s correction for continuity of independence and Bonferroni correction for multiple testing), fold enrichments near <t>TNFα</t> target genes (HAEC RNA-seq, ±10 kb of transcriptional start sites, p value: * <1.0 × 10−5, two-sided Fisher’s exact test with Bonferroni correction), and the top GO terms (GREAT FDR q values, Biological Process Ontology, Supplementary Dataset 3) for the pan-cell (four-cell- type-shared), EC-specific, and HAEC-specific RELA peaks. ↑= upregulated, ↓= downregulated, NC no change (i.e., constitutively expressed). d Profile plots comparing ERG binding signal at pan-cell, EC-specific, and HAEC-specific RELA peaks in HAECs. ERG ChIP-seq signal was generated using published raw data from ref. 15. The plot lines indicate mean RPM ± SEM. Arrows and percentages indicate change in signal after TNFα stimulation. The p values were calculated using two-sided Welch Two Sample t-test (p value: *<0.05). e, Model of NF-κB-ERG dynamics and cofactor squelching showing redistribution of cofactors from EC-specific Mode O regions to pan-cell Mode P regions. Source data are provided in the Source Data file for Fig. 4c, d.
Mouse Brain Tumor Cell Line Kt 98, supplied by ATCC, 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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mouse leydig tumor cell line  (ATCC)
94
ATCC mouse leydig tumor cell line
Fig. 4 Conserved NF-κB binding is shared between cell types near common immune response genes. a Representative genomic regions showing the pan-cell (four-cell-type-shared: between human aortic ECs [HAECs], human umbilical vein ECs [HUVECs], lymphoblastoid cell lines [LCLs], and adipocytes), EC-specific (shared only between HAECs and HUVECs), LCL-specific, and adipocyte-specific RELA peaks. b Stacked bar charts showing the fractions of four-, three-, and two-cell-type-shared and HAEC-specific RELA peaks (one-cell type) that are three-species-conserved, two-species- conserved, or human-specific. c Table showing the top-scoring de novo motifs (MEME-ChIP e values, Supplementary Dataset 4), fold enrichments of RELA binding modes (p value: *<1.0 × 10−6, Chi-squared test with Yates’s correction for continuity of independence and Bonferroni correction for multiple testing), fold enrichments near <t>TNFα</t> target genes (HAEC RNA-seq, ±10 kb of transcriptional start sites, p value: * <1.0 × 10−5, two-sided Fisher’s exact test with Bonferroni correction), and the top GO terms (GREAT FDR q values, Biological Process Ontology, Supplementary Dataset 3) for the pan-cell (four-cell- type-shared), EC-specific, and HAEC-specific RELA peaks. ↑= upregulated, ↓= downregulated, NC no change (i.e., constitutively expressed). d Profile plots comparing ERG binding signal at pan-cell, EC-specific, and HAEC-specific RELA peaks in HAECs. ERG ChIP-seq signal was generated using published raw data from ref. 15. The plot lines indicate mean RPM ± SEM. Arrows and percentages indicate change in signal after TNFα stimulation. The p values were calculated using two-sided Welch Two Sample t-test (p value: *<0.05). e, Model of NF-κB-ERG dynamics and cofactor squelching showing redistribution of cofactors from EC-specific Mode O regions to pan-cell Mode P regions. Source data are provided in the Source Data file for Fig. 4c, d.
Mouse Leydig Tumor Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse mammary tumor cell line mmc  (ATCC)
93
ATCC mouse mammary tumor cell line mmc
(A) TgMMTV-neu mice received three vaccinations with empty vector and each of the seven early tumor antigens. <t>MMC</t> tumor cells were implanted on day 0. (B) TgMMTV-neu mice received three vaccinations with empty vector ● and the five late stage tumor antigens that have human homologs. MMC tumor cells were implanted on day 0 ***p<0.001. (C) Vaccination Schema. <t>(D)MMC</t> <t>syngeneic</t> tumor lysate but not lysate from FVB splenocytes induces an IFN-g T cell immune response in mice vaccinated with the early and late stage tumor antigens. IFN-g secreting cells quantified as precursor frequency (y-axis) for FVB mice (n=5) vaccinated with the plasmids for the early tumor antigens (light gray) or FVB mice (n=5) vaccinated with the plasmids for the late stage antigens (dark gray) as compared to FVB mice (n=5) vaccinated with empty control vector plasmid (white). The positive control is concanavalin A (CONA). * p<0.05 (E) Vaccination with the early and late stage tumor antigens induces a CD8+ T cell immune response. Granzyme B ELISA was performed on the pooled supernatants from the ELISPOT assay. ** p=0.01 * p<0.05
Mouse Mammary Tumor Cell Line Mmc, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse rabbit double antibody  (Boster Bio)
90
Boster Bio mouse rabbit double antibody
(A) TgMMTV-neu mice received three vaccinations with empty vector and each of the seven early tumor antigens. <t>MMC</t> tumor cells were implanted on day 0. (B) TgMMTV-neu mice received three vaccinations with empty vector ● and the five late stage tumor antigens that have human homologs. MMC tumor cells were implanted on day 0 ***p<0.001. (C) Vaccination Schema. <t>(D)MMC</t> <t>syngeneic</t> tumor lysate but not lysate from FVB splenocytes induces an IFN-g T cell immune response in mice vaccinated with the early and late stage tumor antigens. IFN-g secreting cells quantified as precursor frequency (y-axis) for FVB mice (n=5) vaccinated with the plasmids for the early tumor antigens (light gray) or FVB mice (n=5) vaccinated with the plasmids for the late stage antigens (dark gray) as compared to FVB mice (n=5) vaccinated with empty control vector plasmid (white). The positive control is concanavalin A (CONA). * p<0.05 (E) Vaccination with the early and late stage tumor antigens induces a CD8+ T cell immune response. Granzyme B ELISA was performed on the pooled supernatants from the ELISPOT assay. ** p=0.01 * p<0.05
Mouse Rabbit Double Antibody, supplied by Boster Bio, 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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105 mouse ct 26 luc cancer cells  (Genecopoeia)
94
Genecopoeia 105 mouse ct 26 luc cancer cells
(A) TgMMTV-neu mice received three vaccinations with empty vector and each of the seven early tumor antigens. <t>MMC</t> tumor cells were implanted on day 0. (B) TgMMTV-neu mice received three vaccinations with empty vector ● and the five late stage tumor antigens that have human homologs. MMC tumor cells were implanted on day 0 ***p<0.001. (C) Vaccination Schema. <t>(D)MMC</t> <t>syngeneic</t> tumor lysate but not lysate from FVB splenocytes induces an IFN-g T cell immune response in mice vaccinated with the early and late stage tumor antigens. IFN-g secreting cells quantified as precursor frequency (y-axis) for FVB mice (n=5) vaccinated with the plasmids for the early tumor antigens (light gray) or FVB mice (n=5) vaccinated with the plasmids for the late stage antigens (dark gray) as compared to FVB mice (n=5) vaccinated with empty control vector plasmid (white). The positive control is concanavalin A (CONA). * p<0.05 (E) Vaccination with the early and late stage tumor antigens induces a CD8+ T cell immune response. Granzyme B ELISA was performed on the pooled supernatants from the ELISPOT assay. ** p=0.01 * p<0.05
105 Mouse Ct 26 Luc Cancer Cells, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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b16-f10 melanoma cell line  (China Center for Type Culture Collection)
90
China Center for Type Culture Collection b16-f10 melanoma cell line
(A) TgMMTV-neu mice received three vaccinations with empty vector and each of the seven early tumor antigens. <t>MMC</t> tumor cells were implanted on day 0. (B) TgMMTV-neu mice received three vaccinations with empty vector ● and the five late stage tumor antigens that have human homologs. MMC tumor cells were implanted on day 0 ***p<0.001. (C) Vaccination Schema. <t>(D)MMC</t> <t>syngeneic</t> tumor lysate but not lysate from FVB splenocytes induces an IFN-g T cell immune response in mice vaccinated with the early and late stage tumor antigens. IFN-g secreting cells quantified as precursor frequency (y-axis) for FVB mice (n=5) vaccinated with the plasmids for the early tumor antigens (light gray) or FVB mice (n=5) vaccinated with the plasmids for the late stage antigens (dark gray) as compared to FVB mice (n=5) vaccinated with empty control vector plasmid (white). The positive control is concanavalin A (CONA). * p<0.05 (E) Vaccination with the early and late stage tumor antigens induces a CD8+ T cell immune response. Granzyme B ELISA was performed on the pooled supernatants from the ELISPOT assay. ** p=0.01 * p<0.05
B16 F10 Melanoma Cell Line, supplied by China Center for Type Culture Collection, 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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neural stem cell culture and derivation of mouse tumor cell lines  (StemCells Inc)
90
StemCells Inc neural stem cell culture and derivation of mouse tumor cell lines
(A) TgMMTV-neu mice received three vaccinations with empty vector and each of the seven early tumor antigens. <t>MMC</t> tumor cells were implanted on day 0. (B) TgMMTV-neu mice received three vaccinations with empty vector ● and the five late stage tumor antigens that have human homologs. MMC tumor cells were implanted on day 0 ***p<0.001. (C) Vaccination Schema. <t>(D)MMC</t> <t>syngeneic</t> tumor lysate but not lysate from FVB splenocytes induces an IFN-g T cell immune response in mice vaccinated with the early and late stage tumor antigens. IFN-g secreting cells quantified as precursor frequency (y-axis) for FVB mice (n=5) vaccinated with the plasmids for the early tumor antigens (light gray) or FVB mice (n=5) vaccinated with the plasmids for the late stage antigens (dark gray) as compared to FVB mice (n=5) vaccinated with empty control vector plasmid (white). The positive control is concanavalin A (CONA). * p<0.05 (E) Vaccination with the early and late stage tumor antigens induces a CD8+ T cell immune response. Granzyme B ELISA was performed on the pooled supernatants from the ELISPOT assay. ** p=0.01 * p<0.05
Neural Stem Cell Culture And Derivation Of Mouse Tumor Cell Lines, supplied by StemCells Inc, 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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mouse mammary tumor cell line eo771  (Beijing Zhongyuan)
90
Beijing Zhongyuan mouse mammary tumor cell line eo771
(A) TgMMTV-neu mice received three vaccinations with empty vector and each of the seven early tumor antigens. <t>MMC</t> tumor cells were implanted on day 0. (B) TgMMTV-neu mice received three vaccinations with empty vector ● and the five late stage tumor antigens that have human homologs. MMC tumor cells were implanted on day 0 ***p<0.001. (C) Vaccination Schema. <t>(D)MMC</t> <t>syngeneic</t> tumor lysate but not lysate from FVB splenocytes induces an IFN-g T cell immune response in mice vaccinated with the early and late stage tumor antigens. IFN-g secreting cells quantified as precursor frequency (y-axis) for FVB mice (n=5) vaccinated with the plasmids for the early tumor antigens (light gray) or FVB mice (n=5) vaccinated with the plasmids for the late stage antigens (dark gray) as compared to FVB mice (n=5) vaccinated with empty control vector plasmid (white). The positive control is concanavalin A (CONA). * p<0.05 (E) Vaccination with the early and late stage tumor antigens induces a CD8+ T cell immune response. Granzyme B ELISA was performed on the pooled supernatants from the ELISPOT assay. ** p=0.01 * p<0.05
Mouse Mammary Tumor Cell Line Eo771, supplied by Beijing Zhongyuan, 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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Image Search Results


a Relative quantification of Kif5a, Kif5b and Kif5c transcripts by real-time PCR in CD8α + and CD11b + DCs and BMDCs from WT or cKO Kif5b mice. Transcript levels for each sample are expressed relative to Kif5b. Graph shows mean ± S.E.M. ( n = 3). b Contour plots of DCs from the spleen of either WT or cKO Kif5b mice pregated on CD19 − , Gr − , F4/80 − CD11c + , and IA/IE + . The data are representative of three independent experiments. c Absolute numbers of CD11b + DCs and CD8α + DCs in the spleen of WT (blue circles) and cKO Kif5b (red squares) mice. The data are representative of five independent experiments. d – g The efficiency of cross-presentation of different concentration of sOVA and the presentation of OVA peptide SIINFEKL in vitro by CD8α + DCs ( d , g ), CD11b + DCs ( e , g ) and BMDCs ( f , g ) from WT (blue histogram or line) or cKO Kif5b (red histogram or line) mice was measured as IL-2 secretion by OT-I T cells after 16 h of co-culture. Graph shows mean ± S.E.M. ( n = 4). Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. * P < 0.05; ** P < 0.005; *** P < 0.0001.

Journal: Nature Communications

Article Title: Kinesin-1 regulates antigen cross-presentation through the scission of tubulations from early endosomes in dendritic cells

doi: 10.1038/s41467-020-15692-0

Figure Lengend Snippet: a Relative quantification of Kif5a, Kif5b and Kif5c transcripts by real-time PCR in CD8α + and CD11b + DCs and BMDCs from WT or cKO Kif5b mice. Transcript levels for each sample are expressed relative to Kif5b. Graph shows mean ± S.E.M. ( n = 3). b Contour plots of DCs from the spleen of either WT or cKO Kif5b mice pregated on CD19 − , Gr − , F4/80 − CD11c + , and IA/IE + . The data are representative of three independent experiments. c Absolute numbers of CD11b + DCs and CD8α + DCs in the spleen of WT (blue circles) and cKO Kif5b (red squares) mice. The data are representative of five independent experiments. d – g The efficiency of cross-presentation of different concentration of sOVA and the presentation of OVA peptide SIINFEKL in vitro by CD8α + DCs ( d , g ), CD11b + DCs ( e , g ) and BMDCs ( f , g ) from WT (blue histogram or line) or cKO Kif5b (red histogram or line) mice was measured as IL-2 secretion by OT-I T cells after 16 h of co-culture. Graph shows mean ± S.E.M. ( n = 4). Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. * P < 0.05; ** P < 0.005; *** P < 0.0001.

Article Snippet: Anti-CD11c (BV711 (BioLegend), APC (BD Biosciences) PB (Sony)), anti-IA/IE FITC (BD Biosciences), anti-CD8α BV450 (Sony), anti-CD11b (PE (BD Biosciences), APC-CY7 (Sony)), anti-F4/80 PE/Cy7 (Sony), anti-GR1 APC (BD Biosciences), anti-CD19 (PerCP (BD Biosciences), PerCP/Cy5.5 (Sony)), anti-B220 BV650 (BioLegend), anti-CD86 BV650 (Sony), anti-CD40 PE/Cy7 (Sony), mouse monoclonal Kif5b antibody (Biolegend), purified rabbit anti-α-tubulin (Rockland), goat anti-EEA1 antibody (Santa Cruz Biotechnology), rat anti-Lamp1 antibody (Invitrogen), H-2Kb PE antibody (Biolegend), AF6-88.5 clone (Biolegend), rabbit Rab11 antibody (Life Technologies), anti-CD71 PE antibody (Sony), Alexa Fluor-594 donkey anti-rat IgG (H+L) (Life Technologies), Alexa Fluor-555 donkey anti-goat IgG (H+L) (Life Technologies), Alexa Fluor-488 donkey anti-mouse IgG (H+L) (Life Technologies), and Alexa Fluor-555 donkey anti-rabbit IgG (H+L) (Life Technologies) were used.

Techniques: Quantitative Proteomics, Real-time Polymerase Chain Reaction, Concentration Assay, In Vitro, Co-Culture Assay, Comparison

a Tumour growth curves for WT (blue line) or cKO Kif5b (orange line) mice injected subcutaneously with B16-OVA cells and adoptively transferred with OT-I T cells (WT purple line, cKO Kif5b yellow line). Tumour growth was monitored daily, and non-survival was defined as ulceration of the tumour or a mean tumour diameter of 15 mm. The data are quoted as the mean ± SEM from two independent experiments (WT, WT OT1, cKO Kif5b n = 10 mice per group, cKO Kif5b OT1 n = 9 mice). Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. * P < 0.05; ** P < 0.005; *** P < 0.0001. b Survival curves for WT or cKO Kif5b mice, treated as in a . Statistical significance was determined by the log-rank test and by the Gehan–Breslow–Wilcoxon test. * P < 0.05; ** P < 0.005. c Mice were injected with Violet-labelled transgenic OT-I T cells and primed 1 day later with CD11c/P3UOVA (WT blue line or circles, cKO Kif5b red line or circles; WT mice n = 7, cKO Kif5b mice n = 7). The left panel shows representative profiles gated on CD8 + , TCR Vα2 + cells. Statistical analysis of the division index is shown in the right panel: ** P < 0.005 in a two-tailed unpaired Student’s t test. Graph shows mean ± S.E.M.

Journal: Nature Communications

Article Title: Kinesin-1 regulates antigen cross-presentation through the scission of tubulations from early endosomes in dendritic cells

doi: 10.1038/s41467-020-15692-0

Figure Lengend Snippet: a Tumour growth curves for WT (blue line) or cKO Kif5b (orange line) mice injected subcutaneously with B16-OVA cells and adoptively transferred with OT-I T cells (WT purple line, cKO Kif5b yellow line). Tumour growth was monitored daily, and non-survival was defined as ulceration of the tumour or a mean tumour diameter of 15 mm. The data are quoted as the mean ± SEM from two independent experiments (WT, WT OT1, cKO Kif5b n = 10 mice per group, cKO Kif5b OT1 n = 9 mice). Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. * P < 0.05; ** P < 0.005; *** P < 0.0001. b Survival curves for WT or cKO Kif5b mice, treated as in a . Statistical significance was determined by the log-rank test and by the Gehan–Breslow–Wilcoxon test. * P < 0.05; ** P < 0.005. c Mice were injected with Violet-labelled transgenic OT-I T cells and primed 1 day later with CD11c/P3UOVA (WT blue line or circles, cKO Kif5b red line or circles; WT mice n = 7, cKO Kif5b mice n = 7). The left panel shows representative profiles gated on CD8 + , TCR Vα2 + cells. Statistical analysis of the division index is shown in the right panel: ** P < 0.005 in a two-tailed unpaired Student’s t test. Graph shows mean ± S.E.M.

Article Snippet: Anti-CD11c (BV711 (BioLegend), APC (BD Biosciences) PB (Sony)), anti-IA/IE FITC (BD Biosciences), anti-CD8α BV450 (Sony), anti-CD11b (PE (BD Biosciences), APC-CY7 (Sony)), anti-F4/80 PE/Cy7 (Sony), anti-GR1 APC (BD Biosciences), anti-CD19 (PerCP (BD Biosciences), PerCP/Cy5.5 (Sony)), anti-B220 BV650 (BioLegend), anti-CD86 BV650 (Sony), anti-CD40 PE/Cy7 (Sony), mouse monoclonal Kif5b antibody (Biolegend), purified rabbit anti-α-tubulin (Rockland), goat anti-EEA1 antibody (Santa Cruz Biotechnology), rat anti-Lamp1 antibody (Invitrogen), H-2Kb PE antibody (Biolegend), AF6-88.5 clone (Biolegend), rabbit Rab11 antibody (Life Technologies), anti-CD71 PE antibody (Sony), Alexa Fluor-594 donkey anti-rat IgG (H+L) (Life Technologies), Alexa Fluor-555 donkey anti-goat IgG (H+L) (Life Technologies), Alexa Fluor-488 donkey anti-mouse IgG (H+L) (Life Technologies), and Alexa Fluor-555 donkey anti-rabbit IgG (H+L) (Life Technologies) were used.

Techniques: Injection, Comparison, Transgenic Assay, Two Tailed Test

a CD8α + , CD11b + DCs and BMDCs from WT (blue histogram) or cKO Kif5b (red histogram) mice were incubated for 10, 30 or 60 min at 37 °C or for 10 min at 4 °C with Alexa-Flour-647-OVA prior to flow cytometry analysis. The figure shows a histogram that was representative of three independent experiments. b CD8α + , CD11b + DCs and BMDCs from WT (blue line) or cKO Kif5b (red line) mice were incubated for 30 min at 37 °C with DQ-OVA. Next, the cells were washed. DQ-OVA degradation was analysed by flow cytometry after different chase periods. Graphs are representative of four independent experiments. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. *** P < 0.0001. c Degradation of DQ-OVA in BMDCs from WT (DMSO blue line, Conc-A or Cath I purple line) or cKO Kif5b (DMSO orange line, Conc-A or Cath I yellow line) mice pretreated or not with concanamycin A (Conc-A) or a cathepsin inhibitor-I (Cath I). Graphs are representative of four independent experiments. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. ** P < 0.005; *** P < 0.0001. d The pH values in CD8α + , CD11b + DCs and BMDCs from WT (blue line) or cKO Kif5b (red line) mice were determined by FACS after a pulse and different chase periods. Graphs are representative of four independent experiments. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. * P < 0.05; ** P < 0.005; *** P < 0.0001. e Protease activity of CatB/L in early (20 min) or late (120 min) using total cell lysate from BMDCs from WT (blue histogram) or cKO Kif5b (red histogram) mice was measured with a specific fluorescent substrate. Graphs are representative of three independent experiments. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. *** P < 0.0001. a – e Graphs show mean ± S.E.M.

Journal: Nature Communications

Article Title: Kinesin-1 regulates antigen cross-presentation through the scission of tubulations from early endosomes in dendritic cells

doi: 10.1038/s41467-020-15692-0

Figure Lengend Snippet: a CD8α + , CD11b + DCs and BMDCs from WT (blue histogram) or cKO Kif5b (red histogram) mice were incubated for 10, 30 or 60 min at 37 °C or for 10 min at 4 °C with Alexa-Flour-647-OVA prior to flow cytometry analysis. The figure shows a histogram that was representative of three independent experiments. b CD8α + , CD11b + DCs and BMDCs from WT (blue line) or cKO Kif5b (red line) mice were incubated for 30 min at 37 °C with DQ-OVA. Next, the cells were washed. DQ-OVA degradation was analysed by flow cytometry after different chase periods. Graphs are representative of four independent experiments. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. *** P < 0.0001. c Degradation of DQ-OVA in BMDCs from WT (DMSO blue line, Conc-A or Cath I purple line) or cKO Kif5b (DMSO orange line, Conc-A or Cath I yellow line) mice pretreated or not with concanamycin A (Conc-A) or a cathepsin inhibitor-I (Cath I). Graphs are representative of four independent experiments. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. ** P < 0.005; *** P < 0.0001. d The pH values in CD8α + , CD11b + DCs and BMDCs from WT (blue line) or cKO Kif5b (red line) mice were determined by FACS after a pulse and different chase periods. Graphs are representative of four independent experiments. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. * P < 0.05; ** P < 0.005; *** P < 0.0001. e Protease activity of CatB/L in early (20 min) or late (120 min) using total cell lysate from BMDCs from WT (blue histogram) or cKO Kif5b (red histogram) mice was measured with a specific fluorescent substrate. Graphs are representative of three independent experiments. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. *** P < 0.0001. a – e Graphs show mean ± S.E.M.

Article Snippet: Anti-CD11c (BV711 (BioLegend), APC (BD Biosciences) PB (Sony)), anti-IA/IE FITC (BD Biosciences), anti-CD8α BV450 (Sony), anti-CD11b (PE (BD Biosciences), APC-CY7 (Sony)), anti-F4/80 PE/Cy7 (Sony), anti-GR1 APC (BD Biosciences), anti-CD19 (PerCP (BD Biosciences), PerCP/Cy5.5 (Sony)), anti-B220 BV650 (BioLegend), anti-CD86 BV650 (Sony), anti-CD40 PE/Cy7 (Sony), mouse monoclonal Kif5b antibody (Biolegend), purified rabbit anti-α-tubulin (Rockland), goat anti-EEA1 antibody (Santa Cruz Biotechnology), rat anti-Lamp1 antibody (Invitrogen), H-2Kb PE antibody (Biolegend), AF6-88.5 clone (Biolegend), rabbit Rab11 antibody (Life Technologies), anti-CD71 PE antibody (Sony), Alexa Fluor-594 donkey anti-rat IgG (H+L) (Life Technologies), Alexa Fluor-555 donkey anti-goat IgG (H+L) (Life Technologies), Alexa Fluor-488 donkey anti-mouse IgG (H+L) (Life Technologies), and Alexa Fluor-555 donkey anti-rabbit IgG (H+L) (Life Technologies) were used.

Techniques: Incubation, Flow Cytometry, Comparison, Activity Assay

BMDCs from WT or cKO Kif5b mice were incubated for 15, 30 or 60 min with Alexa-Flour-647-OVA and then plated on glass coverslips. The cells were fixed, permeabilized and stained with anti-EEA1, anti-Lamp1 or anti-Rab11 antibodies. The left panel shown representative images for the 15 min time point observed in three independent experiments. Bars: 5 μm. n = 32 cells per condition. Statistical significance (shown in the right panel; WT blue circle, cKO Kif5b red square) was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. ** P < 0.005; *** P < 0.0001. Graphs show mean ± S.E.M.

Journal: Nature Communications

Article Title: Kinesin-1 regulates antigen cross-presentation through the scission of tubulations from early endosomes in dendritic cells

doi: 10.1038/s41467-020-15692-0

Figure Lengend Snippet: BMDCs from WT or cKO Kif5b mice were incubated for 15, 30 or 60 min with Alexa-Flour-647-OVA and then plated on glass coverslips. The cells were fixed, permeabilized and stained with anti-EEA1, anti-Lamp1 or anti-Rab11 antibodies. The left panel shown representative images for the 15 min time point observed in three independent experiments. Bars: 5 μm. n = 32 cells per condition. Statistical significance (shown in the right panel; WT blue circle, cKO Kif5b red square) was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. ** P < 0.005; *** P < 0.0001. Graphs show mean ± S.E.M.

Article Snippet: Anti-CD11c (BV711 (BioLegend), APC (BD Biosciences) PB (Sony)), anti-IA/IE FITC (BD Biosciences), anti-CD8α BV450 (Sony), anti-CD11b (PE (BD Biosciences), APC-CY7 (Sony)), anti-F4/80 PE/Cy7 (Sony), anti-GR1 APC (BD Biosciences), anti-CD19 (PerCP (BD Biosciences), PerCP/Cy5.5 (Sony)), anti-B220 BV650 (BioLegend), anti-CD86 BV650 (Sony), anti-CD40 PE/Cy7 (Sony), mouse monoclonal Kif5b antibody (Biolegend), purified rabbit anti-α-tubulin (Rockland), goat anti-EEA1 antibody (Santa Cruz Biotechnology), rat anti-Lamp1 antibody (Invitrogen), H-2Kb PE antibody (Biolegend), AF6-88.5 clone (Biolegend), rabbit Rab11 antibody (Life Technologies), anti-CD71 PE antibody (Sony), Alexa Fluor-594 donkey anti-rat IgG (H+L) (Life Technologies), Alexa Fluor-555 donkey anti-goat IgG (H+L) (Life Technologies), Alexa Fluor-488 donkey anti-mouse IgG (H+L) (Life Technologies), and Alexa Fluor-555 donkey anti-rabbit IgG (H+L) (Life Technologies) were used.

Techniques: Incubation, Staining, Comparison

a CD8α + , CD11b + DCs and BMDCs from WT (blue line) or cKO Kif5b (red line) mice were labelled for H-2K b and analysed using FACS. The figure shows a FACS histogram that was representative of three independent experiments. b MHC-I recycling ability in CD8α + , CD11b + DCs and BMDCs from WT (blue line) or cKO Kif5b (red line) mice was measured using FACS at the indicated time points. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. ** P < 0.005; *** P < 0.0001. c TrfR (CD71) recycling ability was measured using FACS at the indicated time periods in CD8α + , CD11b + DCs and BMDCs from WT or cKO kif5b mice. Statistical analysis: * P < 0.05; ** P < 0.005; *** P < 0.0001 in the two-way ANOVA and Sidak test’s correction for multiple comparison. d MHC-I recycling in the presence of primaquine in BMDCs from WT or cKO Kif5b mice was measured using FACS at the indicated time points. Statistical analysis: ** P < 0.005; in the two-way ANOVA and Sidak test’s correction for multiple comparison. b – d Graphs are representative of four independent experiments. e BMDCs from WT or cKO kif5b mice were plated on glass coverslips and were fixed, permeabilized and stained with anti-EEA1, anti-Rab11 or anti-MHC-I antibodies. Representative images are shown in the left panel observed in three independent experiments. ( n = 33 cells per condition upper panel, n = 29 cells per condition lower panel) Bars: 5 μm. Statistical analysis (shown in the right panel; WT blue circle, cKO Kif5b red square): ** P < 0.005; *** P < 0.0001 in an two-tailed unpaired Student’s t test. b – e Graphs show mean ± S.E.M.

Journal: Nature Communications

Article Title: Kinesin-1 regulates antigen cross-presentation through the scission of tubulations from early endosomes in dendritic cells

doi: 10.1038/s41467-020-15692-0

Figure Lengend Snippet: a CD8α + , CD11b + DCs and BMDCs from WT (blue line) or cKO Kif5b (red line) mice were labelled for H-2K b and analysed using FACS. The figure shows a FACS histogram that was representative of three independent experiments. b MHC-I recycling ability in CD8α + , CD11b + DCs and BMDCs from WT (blue line) or cKO Kif5b (red line) mice was measured using FACS at the indicated time points. Statistical significance was determined by the two-way ANOVA and Sidak test’s correction for multiple comparison. ** P < 0.005; *** P < 0.0001. c TrfR (CD71) recycling ability was measured using FACS at the indicated time periods in CD8α + , CD11b + DCs and BMDCs from WT or cKO kif5b mice. Statistical analysis: * P < 0.05; ** P < 0.005; *** P < 0.0001 in the two-way ANOVA and Sidak test’s correction for multiple comparison. d MHC-I recycling in the presence of primaquine in BMDCs from WT or cKO Kif5b mice was measured using FACS at the indicated time points. Statistical analysis: ** P < 0.005; in the two-way ANOVA and Sidak test’s correction for multiple comparison. b – d Graphs are representative of four independent experiments. e BMDCs from WT or cKO kif5b mice were plated on glass coverslips and were fixed, permeabilized and stained with anti-EEA1, anti-Rab11 or anti-MHC-I antibodies. Representative images are shown in the left panel observed in three independent experiments. ( n = 33 cells per condition upper panel, n = 29 cells per condition lower panel) Bars: 5 μm. Statistical analysis (shown in the right panel; WT blue circle, cKO Kif5b red square): ** P < 0.005; *** P < 0.0001 in an two-tailed unpaired Student’s t test. b – e Graphs show mean ± S.E.M.

Article Snippet: Anti-CD11c (BV711 (BioLegend), APC (BD Biosciences) PB (Sony)), anti-IA/IE FITC (BD Biosciences), anti-CD8α BV450 (Sony), anti-CD11b (PE (BD Biosciences), APC-CY7 (Sony)), anti-F4/80 PE/Cy7 (Sony), anti-GR1 APC (BD Biosciences), anti-CD19 (PerCP (BD Biosciences), PerCP/Cy5.5 (Sony)), anti-B220 BV650 (BioLegend), anti-CD86 BV650 (Sony), anti-CD40 PE/Cy7 (Sony), mouse monoclonal Kif5b antibody (Biolegend), purified rabbit anti-α-tubulin (Rockland), goat anti-EEA1 antibody (Santa Cruz Biotechnology), rat anti-Lamp1 antibody (Invitrogen), H-2Kb PE antibody (Biolegend), AF6-88.5 clone (Biolegend), rabbit Rab11 antibody (Life Technologies), anti-CD71 PE antibody (Sony), Alexa Fluor-594 donkey anti-rat IgG (H+L) (Life Technologies), Alexa Fluor-555 donkey anti-goat IgG (H+L) (Life Technologies), Alexa Fluor-488 donkey anti-mouse IgG (H+L) (Life Technologies), and Alexa Fluor-555 donkey anti-rabbit IgG (H+L) (Life Technologies) were used.

Techniques: Comparison, Staining, Two Tailed Test

a Confocal microscopy of BMDCs from WT or cKO Kif5b mice stained with anti-tubulin and anti-Kif5b. Bars: 5 μm. The indicated box is shown at a higher magnification in the inset. Bars: 2 μm. b Confocal microscopy of WT BMDCs stained with anti-EEA1 and anti-Kif5b. The intersection between the two stainings is shown in white. The indicated boxes are shown at higher magnification in the insets. Bars: 2 μm. c A schematic representation of early endosome WGA labelling. d WT and cKO Kif5b BMDCs were labelled with WGA-488 and stained with anti-EEA1. Bars: 2 μm (left panel). Quantification of the colocalization between WGA-488 and EEA1-555 (right panel; WT blue histogram, cKO Kif5b red histogram; n = 7 cells per condition). All images of single cells are representative of >100 cells observed in three independent experiments ( a – d ). e Spinning disk video microscopy was performed on WT and cKO Kif5b BMDCs labelled with WGA-488. Representative series of images are shown every 25 s. Bars: 2 μm. Images are representative of four independent experiments. See also Supplementary Movies and . f The number of tubulations unable to detach per cell during the 5 min acquisition. (WT blue circles, cKO Kif5b red squares; n = 20 cells per condition). Statistical analysis: *** P < 0.0001 in a two-tailed unpaired Student’s t test. g The number of WGA-488-positive vesicular structures >1 μm in size was measured in WT (blue line) and cKO Kif5b (red line) BMDCs during the 5 min acquisition ( n = 13 cells per condition). Statistical analysis: *** P < 0.0001 in a two-way ANOVA and Sidak test’s correction for multiple comparison. h Schematic representation of the WGA labelling protocol (upper panel). Spinning disk video microscopy was performed on WT and cKO Kif5b BMDCs labelled with WGA-555 and WGA-488. Representative series of images observed in three independent experiments are shown at the indicated time periods (lower panel). Bars: 2 μm. i n = 10 cells per condition. Statistical analysis of the experiment in (WT blue histogram, cKO Kif5b red histogram) ( h ); *** P < 0.0001 in a two-way ANOVA and Sidak test’s correction for multiple comparison. d – i Graphs show mean ± S.E.M.

Journal: Nature Communications

Article Title: Kinesin-1 regulates antigen cross-presentation through the scission of tubulations from early endosomes in dendritic cells

doi: 10.1038/s41467-020-15692-0

Figure Lengend Snippet: a Confocal microscopy of BMDCs from WT or cKO Kif5b mice stained with anti-tubulin and anti-Kif5b. Bars: 5 μm. The indicated box is shown at a higher magnification in the inset. Bars: 2 μm. b Confocal microscopy of WT BMDCs stained with anti-EEA1 and anti-Kif5b. The intersection between the two stainings is shown in white. The indicated boxes are shown at higher magnification in the insets. Bars: 2 μm. c A schematic representation of early endosome WGA labelling. d WT and cKO Kif5b BMDCs were labelled with WGA-488 and stained with anti-EEA1. Bars: 2 μm (left panel). Quantification of the colocalization between WGA-488 and EEA1-555 (right panel; WT blue histogram, cKO Kif5b red histogram; n = 7 cells per condition). All images of single cells are representative of >100 cells observed in three independent experiments ( a – d ). e Spinning disk video microscopy was performed on WT and cKO Kif5b BMDCs labelled with WGA-488. Representative series of images are shown every 25 s. Bars: 2 μm. Images are representative of four independent experiments. See also Supplementary Movies and . f The number of tubulations unable to detach per cell during the 5 min acquisition. (WT blue circles, cKO Kif5b red squares; n = 20 cells per condition). Statistical analysis: *** P < 0.0001 in a two-tailed unpaired Student’s t test. g The number of WGA-488-positive vesicular structures >1 μm in size was measured in WT (blue line) and cKO Kif5b (red line) BMDCs during the 5 min acquisition ( n = 13 cells per condition). Statistical analysis: *** P < 0.0001 in a two-way ANOVA and Sidak test’s correction for multiple comparison. h Schematic representation of the WGA labelling protocol (upper panel). Spinning disk video microscopy was performed on WT and cKO Kif5b BMDCs labelled with WGA-555 and WGA-488. Representative series of images observed in three independent experiments are shown at the indicated time periods (lower panel). Bars: 2 μm. i n = 10 cells per condition. Statistical analysis of the experiment in (WT blue histogram, cKO Kif5b red histogram) ( h ); *** P < 0.0001 in a two-way ANOVA and Sidak test’s correction for multiple comparison. d – i Graphs show mean ± S.E.M.

Article Snippet: Anti-CD11c (BV711 (BioLegend), APC (BD Biosciences) PB (Sony)), anti-IA/IE FITC (BD Biosciences), anti-CD8α BV450 (Sony), anti-CD11b (PE (BD Biosciences), APC-CY7 (Sony)), anti-F4/80 PE/Cy7 (Sony), anti-GR1 APC (BD Biosciences), anti-CD19 (PerCP (BD Biosciences), PerCP/Cy5.5 (Sony)), anti-B220 BV650 (BioLegend), anti-CD86 BV650 (Sony), anti-CD40 PE/Cy7 (Sony), mouse monoclonal Kif5b antibody (Biolegend), purified rabbit anti-α-tubulin (Rockland), goat anti-EEA1 antibody (Santa Cruz Biotechnology), rat anti-Lamp1 antibody (Invitrogen), H-2Kb PE antibody (Biolegend), AF6-88.5 clone (Biolegend), rabbit Rab11 antibody (Life Technologies), anti-CD71 PE antibody (Sony), Alexa Fluor-594 donkey anti-rat IgG (H+L) (Life Technologies), Alexa Fluor-555 donkey anti-goat IgG (H+L) (Life Technologies), Alexa Fluor-488 donkey anti-mouse IgG (H+L) (Life Technologies), and Alexa Fluor-555 donkey anti-rabbit IgG (H+L) (Life Technologies) were used.

Techniques: Confocal Microscopy, Staining, Microscopy, Two Tailed Test, Comparison

Fig. 4 Conserved NF-κB binding is shared between cell types near common immune response genes. a Representative genomic regions showing the pan-cell (four-cell-type-shared: between human aortic ECs [HAECs], human umbilical vein ECs [HUVECs], lymphoblastoid cell lines [LCLs], and adipocytes), EC-specific (shared only between HAECs and HUVECs), LCL-specific, and adipocyte-specific RELA peaks. b Stacked bar charts showing the fractions of four-, three-, and two-cell-type-shared and HAEC-specific RELA peaks (one-cell type) that are three-species-conserved, two-species- conserved, or human-specific. c Table showing the top-scoring de novo motifs (MEME-ChIP e values, Supplementary Dataset 4), fold enrichments of RELA binding modes (p value: *<1.0 × 10−6, Chi-squared test with Yates’s correction for continuity of independence and Bonferroni correction for multiple testing), fold enrichments near TNFα target genes (HAEC RNA-seq, ±10 kb of transcriptional start sites, p value: * <1.0 × 10−5, two-sided Fisher’s exact test with Bonferroni correction), and the top GO terms (GREAT FDR q values, Biological Process Ontology, Supplementary Dataset 3) for the pan-cell (four-cell- type-shared), EC-specific, and HAEC-specific RELA peaks. ↑= upregulated, ↓= downregulated, NC no change (i.e., constitutively expressed). d Profile plots comparing ERG binding signal at pan-cell, EC-specific, and HAEC-specific RELA peaks in HAECs. ERG ChIP-seq signal was generated using published raw data from ref. 15. The plot lines indicate mean RPM ± SEM. Arrows and percentages indicate change in signal after TNFα stimulation. The p values were calculated using two-sided Welch Two Sample t-test (p value: *<0.05). e, Model of NF-κB-ERG dynamics and cofactor squelching showing redistribution of cofactors from EC-specific Mode O regions to pan-cell Mode P regions. Source data are provided in the Source Data file for Fig. 4c, d.

Journal: Nature communications

Article Title: Conserved regulatory logic at accessible and inaccessible chromatin during the acute inflammatory response in mammals.

doi: 10.1038/s41467-020-20765-1

Figure Lengend Snippet: Fig. 4 Conserved NF-κB binding is shared between cell types near common immune response genes. a Representative genomic regions showing the pan-cell (four-cell-type-shared: between human aortic ECs [HAECs], human umbilical vein ECs [HUVECs], lymphoblastoid cell lines [LCLs], and adipocytes), EC-specific (shared only between HAECs and HUVECs), LCL-specific, and adipocyte-specific RELA peaks. b Stacked bar charts showing the fractions of four-, three-, and two-cell-type-shared and HAEC-specific RELA peaks (one-cell type) that are three-species-conserved, two-species- conserved, or human-specific. c Table showing the top-scoring de novo motifs (MEME-ChIP e values, Supplementary Dataset 4), fold enrichments of RELA binding modes (p value: *<1.0 × 10−6, Chi-squared test with Yates’s correction for continuity of independence and Bonferroni correction for multiple testing), fold enrichments near TNFα target genes (HAEC RNA-seq, ±10 kb of transcriptional start sites, p value: * <1.0 × 10−5, two-sided Fisher’s exact test with Bonferroni correction), and the top GO terms (GREAT FDR q values, Biological Process Ontology, Supplementary Dataset 3) for the pan-cell (four-cell- type-shared), EC-specific, and HAEC-specific RELA peaks. ↑= upregulated, ↓= downregulated, NC no change (i.e., constitutively expressed). d Profile plots comparing ERG binding signal at pan-cell, EC-specific, and HAEC-specific RELA peaks in HAECs. ERG ChIP-seq signal was generated using published raw data from ref. 15. The plot lines indicate mean RPM ± SEM. Arrows and percentages indicate change in signal after TNFα stimulation. The p values were calculated using two-sided Welch Two Sample t-test (p value: *<0.05). e, Model of NF-κB-ERG dynamics and cofactor squelching showing redistribution of cofactors from EC-specific Mode O regions to pan-cell Mode P regions. Source data are provided in the Source Data file for Fig. 4c, d.

Article Snippet: To induce the acute pro-inflammatory response, HAECs (#1487 and #2139) were treated with 10-ng/mL recombinant human TNFα (Cell Applications, cat# RP1111-50), MAECs (#A092913T2MP and #B092913T2MP) were treated with 10-ng/mL recombinant mouse TNFα (Cell Applications, cat# RP2031-20), and BAECs (#1165 and #1190) were treated with 10-ng/mL recombinant bovine TNFα (R&D Systems, cat# 2279-BT-025) for 45 min in basal Endothelial Cell Growth Media MV2 (PromoCell) without supplements.

Techniques: Binding Assay, RNA Sequencing, ChIP-sequencing, Generated

(A) TgMMTV-neu mice received three vaccinations with empty vector and each of the seven early tumor antigens. MMC tumor cells were implanted on day 0. (B) TgMMTV-neu mice received three vaccinations with empty vector ● and the five late stage tumor antigens that have human homologs. MMC tumor cells were implanted on day 0 ***p<0.001. (C) Vaccination Schema. (D)MMC syngeneic tumor lysate but not lysate from FVB splenocytes induces an IFN-g T cell immune response in mice vaccinated with the early and late stage tumor antigens. IFN-g secreting cells quantified as precursor frequency (y-axis) for FVB mice (n=5) vaccinated with the plasmids for the early tumor antigens (light gray) or FVB mice (n=5) vaccinated with the plasmids for the late stage antigens (dark gray) as compared to FVB mice (n=5) vaccinated with empty control vector plasmid (white). The positive control is concanavalin A (CONA). * p<0.05 (E) Vaccination with the early and late stage tumor antigens induces a CD8+ T cell immune response. Granzyme B ELISA was performed on the pooled supernatants from the ELISPOT assay. ** p=0.01 * p<0.05

Journal: Vaccine

Article Title: Tumor-associated antigens identified early in mouse mammary tumor development can be effective vaccine targets.

doi: 10.1016/j.vaccine.2019.05.024

Figure Lengend Snippet: (A) TgMMTV-neu mice received three vaccinations with empty vector and each of the seven early tumor antigens. MMC tumor cells were implanted on day 0. (B) TgMMTV-neu mice received three vaccinations with empty vector ● and the five late stage tumor antigens that have human homologs. MMC tumor cells were implanted on day 0 ***p<0.001. (C) Vaccination Schema. (D)MMC syngeneic tumor lysate but not lysate from FVB splenocytes induces an IFN-g T cell immune response in mice vaccinated with the early and late stage tumor antigens. IFN-g secreting cells quantified as precursor frequency (y-axis) for FVB mice (n=5) vaccinated with the plasmids for the early tumor antigens (light gray) or FVB mice (n=5) vaccinated with the plasmids for the late stage antigens (dark gray) as compared to FVB mice (n=5) vaccinated with empty control vector plasmid (white). The positive control is concanavalin A (CONA). * p<0.05 (E) Vaccination with the early and late stage tumor antigens induces a CD8+ T cell immune response. Granzyme B ELISA was performed on the pooled supernatants from the ELISPOT assay. ** p=0.01 * p<0.05

Article Snippet: The syngeneic mouse mammary tumor cell line MMC and human cell lines MCF10F (non-malignant breast), HCC1500 (ER positive HER2 negative), HCC70 (ER negative HER2 negative), and SKBR3 (ER negative HER2 positive) were seeded in two 96 well plates (ATCC, Manassas VA).

Techniques: Plasmid Preparation, Control, Positive Control, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Using four pooled siRNA for each target in the syngeneic MMC mouse tumor cell line, grey bars are the early stage antigens measuring (A) survival and (B) caspase 3/7 activity as compared to cells transfected with liposomes but no siRNA (control). For the late stage tumor antigens, the grey bars show the late stage tumor antigens measuring (C) survival and (D) caspase 3/7 activity as compared to cells transfected with liposomes but no siRNA (control). White bars include liposome transfected cells (control), non-targeting control siRNA transfected cells (negative control), and untransfected cells (untx). Apoptosis inducing siRNA transfected cells is the black bar (positive control) ** p<0.01 **** p<0.0001 as compared to control.

Journal: Vaccine

Article Title: Tumor-associated antigens identified early in mouse mammary tumor development can be effective vaccine targets.

doi: 10.1016/j.vaccine.2019.05.024

Figure Lengend Snippet: Using four pooled siRNA for each target in the syngeneic MMC mouse tumor cell line, grey bars are the early stage antigens measuring (A) survival and (B) caspase 3/7 activity as compared to cells transfected with liposomes but no siRNA (control). For the late stage tumor antigens, the grey bars show the late stage tumor antigens measuring (C) survival and (D) caspase 3/7 activity as compared to cells transfected with liposomes but no siRNA (control). White bars include liposome transfected cells (control), non-targeting control siRNA transfected cells (negative control), and untransfected cells (untx). Apoptosis inducing siRNA transfected cells is the black bar (positive control) ** p<0.01 **** p<0.0001 as compared to control.

Article Snippet: The syngeneic mouse mammary tumor cell line MMC and human cell lines MCF10F (non-malignant breast), HCC1500 (ER positive HER2 negative), HCC70 (ER negative HER2 negative), and SKBR3 (ER negative HER2 positive) were seeded in two 96 well plates (ATCC, Manassas VA).

Techniques: Activity Assay, Transfection, Liposomes, Control, Negative Control, Positive Control