Journal: Cell

Article Title: Transcriptional Basis of Mouse and Human Dendritic Cell Heterogeneity

doi: 10.1016/j.cell.2019.09.035

Figure Lengend Snippet: Single-Cell Survey Reveals Heterogeneity of cDC2s with Two Subsets Delineated by Expression of T-Bet (A) Representative contour plot showing gating strategy for splenic DCs in Tbx21 RFP-Cre mice. DCs defined as Lin(CD3,CD19,CD49b,Siglec-F) – Ly6C – CD64 – CD11c + MHCII + . (B) Frequency of T-bet + cDC2s across tissues. Each circle represents one mouse. In the peripheral and mesenteric LN (PLN and MLN), migratory DCs were defined as MHCII hi CD11c int and resident DCs as MHCII int CD11c hi . Error bars represent mean ± SEM. (C) Analysis of RFP + and YFP + splenic cDC2s from Tbx21 RFP-CreERT2 Rosa26 YFP mice, 3 days post tamoxifen gavage. (D) Percent RFP + and YFP + of cDC2 cells. Percent RFP + of YFP + cDC2s at indicated time points post tamoxifen gavage (right). Error bars represent mean ± SEM; n = 3–4 mice per time point. (E) t-SNE embedding of 4,464 DCs. Colors indicate unsupervised clustering by Phenograph (left panel) or classification based on expression of canonical markers (right panel). (F) Expression of canonical DC markers across the transcriptionally defined DC clusters from (E). (G) Proportion of T-bet (RFP + ) cells in each cell cluster identified in (D). (H) Violin plot showing expression of the cell-cycle signature across the DC clusters from (E). (I) Similarity of bulk T-bet – cDC2s, T-bet + cDC2, and cDC1 transcriptomes to the reference single-cell DC clusters (E). Colors represent the correlation coefficient between the cell population identified in the row label and the DC cluster identified by the column label. See also and .

Article Snippet: Anti-mouse CD3ε (PE-Cy7) , Tonbo Biosciences , Cat#60-0031; RRID: AB_2621824 ; Clone 145-2C11.

Techniques: Expressing

Journal: Cell

Article Title: Transcriptional Basis of Mouse and Human Dendritic Cell Heterogeneity

doi: 10.1016/j.cell.2019.09.035

Figure Lengend Snippet: Single-Cell Survey Reveals Heterogeneity of cDC2s, Related to Figure 1 A. Representative histogram showing expression of T-bet (RFP) in splenic cells from Tbx21 RFP-cre mice. (B). Expression of T-bet in CD11b + XCR1 + DCs from the intestinal lamina propria. Data representative of > 5 independent experiments, with at least 3 mice per experiment. (C). Expression of T-bet in splenic myeloid cells. Cells were defined as: (i) Ly-6C hi monocytes (Lin – Ly6C + Ly6G – CD11b + CX3CR1 + ); neutrophils (Lin – Ly6C + Ly6G + ); macrophages (Lin – CD64 + Ly6C – ). Lineages (Lin) were defined as: CD3e, CD90.2, CD19, CD49b and Siglec F. Each circle represents an individual mouse, error bars represent mean ± SEM. (D). Left: Gating strategy for single-cell sorting. DCs were defined as Lin(CD3, CD19, CD90) – Ly6C – CD64 – CD11c + MHCII + . Two populations were sampled: RFP + DCs and RFP – DCs (encompassing XCR1 + cDC1s, CD11b + RFP – and CD11b – XCR1 – DCs). Right: Post-sort purity of RFP + and RFP – cells. Contaminating population of Ly6C + cells identifiable on post-sort purity (lower panel). (E). Similarity of splenic CD11c + MHCII + cells to reference myeloid cells (ImmGen Consortium) Colors represent the Pearson correlation between the mean gene expression from the dendritic cell cluster in the rows and the bulk reference transcriptome in the columns. (F). Top 20 positive and negative gene loadings of PC1 for T-bet + cDC2 clusters after cell-cycle correction (left panel). Scatterplot of PC1 and PC2 for T-bet + cDC2 clusters after cell-cycle correction (right panel).

Article Snippet: Anti-mouse CD3ε (PE-Cy7) , Tonbo Biosciences , Cat#60-0031; RRID: AB_2621824 ; Clone 145-2C11.

Techniques: Expressing, FACS, Gene Expression

Journal: Cell

Article Title: Transcriptional Basis of Mouse and Human Dendritic Cell Heterogeneity

doi: 10.1016/j.cell.2019.09.035

Figure Lengend Snippet: Environmental Cues Drive Distinct DC2 Differentiation Pathways within the Spleen, Related to Figure 5 (A). Gating strategy for the identification of DC progenitors in the bone marrow (BM) (B). Palantir pseudo-time analysis of differentiation potential and branch probabilities from the Siglec-H + pre-DC state to T-bet + cDC2 and T-bet – cDC2 terminal states. (C). Plots showing Palantir differentiation potential (y axis) along Palantir pseudo-time (x axis) for Siglec-H + DC and T-bet + cDC2s (top) or Siglec-H + DC and T-bet – cDC2 clusters (bottom) (D). Plots showing the top two diffusion component embeddings for Siglec-H + DC and T-bet + cDC2 clusters (top) or Siglec-H + DC and Tbet – cDC2 clusters (bottom). Black arrow indicates Siglec-H + DC cluster cells adjacent to cells from the proliferative T-bet + cDC2 clusters 6 and 8. (E). Top panel: plots showing probability of each cell being within 20 nearest neighbors of randomly sampled shortest paths from the Siglec-H + DC to the indicated end points. Middle panel: plots showing the proportion of cells belonging to Siglec-H + DC, T-bet + cDC2, or T-bet – cDC2 from 20 nearest neighbors of randomly sampled shortest paths. Bottom: plots showing diffusion distance step sizes for each step along the indicated shortest paths (bottom panel). Colors illustrate cluster membership. (F). Graph showing AUC (x axis) for genes differentially expressed between Siglec-H + DC cluster (cluster 11) and all other cDC2 clusters. EMD on the y axis. Dashed lines represents μ EMD ± 3σ EMD . (G). Gating strategy for FACS-isolation of MHCII + ILC3s: Lin = CD3, CD19, CD49b, Siglec-F. (H). Heatmap reports scaled expression of 3550 differentially expressed genes (log 2 FC > 1, FDR < 0.01) between ILC3s and Rorγt fm cDC2s. Selected genes listed to the right. (I). Representative flow cytometric analysis of phenotypes of splenic progeny from Tbx21 RFP-cre CD45.2 + Ly6C − CD64 – MHCII + CD11c + Siglec-H + pre-DCs adoptively transferred into sub-lethally irradiated CD45.1 recipient mice 7 days earlier (data from one experiment with n = 3). J. Sort purified T-bet + or T-bet – cDC2 were cultured for 24hrs in the presence of LPS, CpG, TNF-α or IFN−γ. Representative overlay histogram showing the expression of RFP(T-bet) at 24hrs. Data representative of 2 (TNF-α) or 4 (all other cytokines/TLR agonists) independent experiments, n = 2-3.

Article Snippet: Anti-mouse CD3ε (PE-Cy7) , Tonbo Biosciences , Cat#60-0031; RRID: AB_2621824 ; Clone 145-2C11.

Techniques: Diffusion-based Assay, Isolation, Expressing, Irradiation, Purification, Cell Culture

Journal: Cell

Article Title: Transcriptional Basis of Mouse and Human Dendritic Cell Heterogeneity

doi: 10.1016/j.cell.2019.09.035

Figure Lengend Snippet: Human DC Heterogeneity, Related to Figure 7 (A). Violin plots showing expression distribution of mouse DC subset marker genes across human peripheral blood DC and monocyte clusters identified in Villani et al. (2017) . (B). Representative flow cytometric analysis of mouse peripheral blood cDC2s showing absence of T-bet (RFP) + cDC2s. (C). Gating strategy for FACS-isolation of human spleen DCs for scRNA-seq. DCs were defined as live, LIN(CD3,CD56,CD19) − CD14 – CD11C + HLA-DR + . (D). Representative flow cytometry analysis of human spleen cDC2s gated as Lin(CD3,CD56,CD19) – CD14 – CD11c + HLA-DR + CD123 – XCR1 – CLEC4A + cells. Left panel: cell surface expression of CD1c and CLEC10A by cDC2s. Right panel: overlay of CLEC10A + and CLEC10A – cDC2s distinguished by differential expression of CLEC4A and FcεR1a. Summary bar graphs show frequency of CD1C + CLEC10A + and CD1C + CLEC10A – cDC2s as a percentage of cDC2s (n = 4 individuals). (E). t -SNE embedding of 9,315 FACS-isolated CD45 + immune cells from two melanoma tumors. Colors indicate unsupervised clustering by Phenograph (left panel) or classification based on expression of canonical markers and correlations with bulk RNA-seq data (right panel). Each dot represents an individual cell. (F). Pearson correlations between cluster centroids in (F) and bulk RNA-seq data from purified immune populations ( Jeffrey et al., 2006 , Novershtern et al., 2011 ) (G). t-SNE map of 2,122 myeloid cells identified in (F). Colors indicate patient sample (left) or unsupervised clustering by Phenograph (right panel). Each dot represents an individual cell. (H). Heatmap of normalized, log transformed and MAGIC imputed expression of top 20 differentially expressed genes, defined by the highest earth mover’s distance (EMD), per Phenograph cluster in E. The colored bar at the top of the heatmap shows assignment of cells to clusters labeled in F, right panel. (I). t-SNE map of human melanoma myeloid cells (H) colored by imputed expression of labeled genes.

Article Snippet: Anti-mouse CD3ε (PE-Cy7) , Tonbo Biosciences , Cat#60-0031; RRID: AB_2621824 ; Clone 145-2C11.

Techniques: Expressing, Marker, Isolation, Flow Cytometry, Quantitative Proteomics, RNA Sequencing, Purification, Transformation Assay, Labeling

Journal: Cell

Article Title: Transcriptional Basis of Mouse and Human Dendritic Cell Heterogeneity

doi: 10.1016/j.cell.2019.09.035

Figure Lengend Snippet:

Article Snippet: Anti-mouse CD3ε (PE-Cy7) , Tonbo Biosciences , Cat#60-0031; RRID: AB_2621824 ; Clone 145-2C11.

Techniques: Recombinant, Staining, Multiplex Assay, Cell Isolation, Gene Expression, Software

Journal: Nature Communications

Article Title: PI3Kγ signaling controls trafficking of CD8 + T cells between lymphoid and non-lymphoid organs and drives hypertension in a murine model

doi: 10.1038/s41467-025-61009-4

Figure Lengend Snippet: a Schematics of the experiment: CD8 + T lymphocytes were purified from total splenocytes from PI3Kγ +/+ , PI3Kγ −/− , and PI3Kγ KD/KD mice receiving angiotensin II or vehicle, and were plated on wells coated with anti-CD3 and anti-CD28 antibodies. PIP3 production was analyzed by immunocytochemistry, and Akt phosphorylation at Thr308 and Ser473 sites by western blot. b , c Immunocytochemistry representative images ( b ) of PIP3 (green) expression on CD8 + T cells (red); DAPI (blue) was used to identify nuclei. Images were acquired by confocal microscopy at 40X magnification (scale bar = 20 µm). c Quantitative analysis of the % of PIP3 + CD8 + T cells from the spleen of n = 3 PI3Kγ +/+ Veh, n = 3 PI3Kγ +/+ AngII, n = 3 PI3Kγ −/− Veh, n = 3 PI3Kγ −/− AngII, n = 3 PI3Kγ KD/KD Veh and n = 3 PI3Kγ KD/KD AngII mice. Representative western blot images ( d , f ) and quantitative analysis ( e, g ) of Akt phosphorylation of Thr308 ( d , e ) and Ser473 ( f , g ) in CD8 + T cells isolated from the spleen of n = 6 PI3Kγ +/+ Veh, n = 6 PI3Kγ +/+ AngII, n = 5 PI3Kγ −/− Veh, n = 5 PI3Kγ −/− AngII, n = 5 PI3Kγ KD/KD Veh and n = 5 PI3Kγ KD/KD AngII mice. Quantitative analysis ( e , g ) as fold changes of pThr308 Akt ( e ) and pSer473 ( g ), normalized over β-actin levels, are shown for angiotensin II treated mice as compared to vehicle controls. All data are expressed as mean ± SEM. Statistical analysis was performed by Two-way ANOVA and Tukey’s correction for multiple comparisons in c , e , g . N represents biologically independent samples. Source data are provided as a file. Schematic in a is Created in BioRender. Perrotta, M. (2025) https://BioRender.com/1qw2qst .

Article Snippet: In the ex-vivo co-culture experiments of MRA and conditioned culture medium of CD8 + T lymphocytes, the cells were isolated from total splenocytes as described above and cultured 5 * 10 5 cells/well at a density of about 7 * 10 5 /cm 2 for 72 h at 37 °C, 5% CO 2 in TexMACS medium (Miltenyi Biotec, 130-097-196) with 10% Dialyzed FBS (Gibco 26400-044), 1% Penicillin-Streptomycin (Gibco, 15140-122), 55 μM β-mercaptoethanol (Gibco, 31350-010) in 48 well pre-coated with CD3ε antibody (pure-functional grade, Miltenyi Biotec, 130-092-973) and then stimulated with CD28 antibody (pure-functional grade, Miltenyi Biotec, 130-093-182).

Techniques: Purification, Immunocytochemistry, Phospho-proteomics, Western Blot, Expressing, Confocal Microscopy, Isolation

Journal: Nature Communications

Article Title: PI3Kγ signaling controls trafficking of CD8 + T cells between lymphoid and non-lymphoid organs and drives hypertension in a murine model

doi: 10.1038/s41467-025-61009-4

Figure Lengend Snippet: a Immunohistochemistry of CD3 + (magenta) and B220 + (green) lymphocytes to analyze the white pulp area in the spleen of PI3Kγ +/+ , PI3Kγ −/− and PI3Kγ KD/KD mice after in vivo administration of angiotensin II or vehicle as control. Nuclei are visualized in blue by DAPI. Images were acquired by confocal microscopy at 10X magnification (scale bar = 100 µm). b Quantitative analysis of the white pulp area marked by CD3 + T cells. n = 6 PI3Kγ +/+ Veh, n = 6 PI3Kγ +/+ AngII, n = 5 PI3Kγ −/− Veh, n = 7 PI3Kγ −/− AngII; n = 7 PI3Kγ KD/KD Veh and n = 5 PI3Kγ KD/KD AngII mice. c Analysis of the myogenic tone of mesenteric resistance arteries (MRA) dissected from naïve WT mice ex vivo co-cultured with CD8 + T cells purified from the spleen of PI3Kγ +/+ and PI3Kγ −/− mice stimulated in vivo for 3 days with angiotensin II or vehicle. n = 4 CD8 Veh ; n = 5 CD8 AngII-PI3Kγ+/+ and n = 5 CD8 AngII-PI3Kγ−/− . Representative plots of flow cytometry ( d ) and quantitative analysis ( e – h ) of total CD8+ T cells ( e ), naïve ( f ), effector ( g ) and resident memory ( h ) CD8+ T cells in kidneys dissected from PI3Kγ +/+ , PI3Kγ −/− and PI3Kγ KD/KD mice after in vivo administration of angiotensin II or vehicle for 28 days. n = 5 PI3Kγ +/+ Veh, n = 7 PI3Kγ +/+ AngII, n = 6 PI3Kγ −/− Veh, n = 8 PI3Kγ −/− AngII, n = 6 PI3Kγ KD/KD Veh and n = 7 PI3Kγ KD/KD AngII. All data are expressed as mean ± SEM. Statistical analysis was performed by Two-way ANOVA, followed by Tukey’s correction for multiple comparisons in b , e , f , g , h ; One-way ANOVA for repeated measures, followed by Tukey’s correction for multiple comparisons in c. N represents biologically independent samples. Source data are provided as a file. Schematics in a , c , d were created in BioRender. Perrotta M (2025) https://BioRender.com/1f77gjq , https://BioRender.com/fbmd4r3 , https://BioRender.com/g0be7ts .

Article Snippet: In the ex-vivo co-culture experiments of MRA and conditioned culture medium of CD8 + T lymphocytes, the cells were isolated from total splenocytes as described above and cultured 5 * 10 5 cells/well at a density of about 7 * 10 5 /cm 2 for 72 h at 37 °C, 5% CO 2 in TexMACS medium (Miltenyi Biotec, 130-097-196) with 10% Dialyzed FBS (Gibco 26400-044), 1% Penicillin-Streptomycin (Gibco, 15140-122), 55 μM β-mercaptoethanol (Gibco, 31350-010) in 48 well pre-coated with CD3ε antibody (pure-functional grade, Miltenyi Biotec, 130-092-973) and then stimulated with CD28 antibody (pure-functional grade, Miltenyi Biotec, 130-093-182).

Techniques: Immunohistochemistry, In Vivo, Control, Confocal Microscopy, Ex Vivo, Cell Culture, Purification, Flow Cytometry

Journal: Nature Communications

Article Title: PI3Kγ signaling controls trafficking of CD8 + T cells between lymphoid and non-lymphoid organs and drives hypertension in a murine model

doi: 10.1038/s41467-025-61009-4

Figure Lengend Snippet: a , b Immunocytochemistry representation ( b ) of PIP3 + (green) expression in CD8 + (red) T cells isolated from the spleen of PI3Kγ +/+ and PI3Kγ CX/CX mice and plated onto anti-CD3 and anti-CD28 coated wells. Nuclei are stained by DAPI and visualized in blue. b Quantitative analysis of the % of PIP3 + CD8 + T cells was performed in n = 3 PI3Kγ +/+ and n = 4 PI3Kγ CX/CX mice. Images were acquired by confocal microscopy at 40X magnification (scale bar = 20 µm). c – f Western blot analysis of Akt phosphorylation in Thr308 (c) and Ser473 ( e ) in CD8 + T cells isolated from the spleens of PI3Kγ +/+ and PI3Kγ CX/CX mice. Representative blot ( c , e ) and quantitative analysis of phosphorylated Akt at Thr308 ( d ) and Ser473 ( f ) as fold changes over β-actin levels, in n = 6 PI3Kγ +/+ and PI3Kγ CX/CX mice in d , and n = 8 PI3Kγ +/+ and PI3Kγ CX/CX mice in ( f ). Metabolic-flux analysis by the Seahorse XF Analyzer was used to measure the mitochondrial respiration and the aerobic glycolysis by the oxygen-consumption rate (OCR) ( g – j ) and extracellular acidification rate (ECAR) ( k – m ) under basal conditions and after drug-induced mitochondrial inhibition in CD8 + T cells purified from the spleen of PI3Kγ CX/CX and PI3Kγ +/+ mice. For g – m , n = 9 PI3Kγ +/+ and n = 8 PI3Kγ CX/CX animals were analyzed and measured in triplicates. All data are expressed as mean ± SEM. Statistical analysis was performed by unpaired Mann–Whitney test in b ; two-sided unpaired T -test in d , f , h , i , j , l , m . N represents biologically independent samples. Source data are provided as a file.

Article Snippet: In the ex-vivo co-culture experiments of MRA and conditioned culture medium of CD8 + T lymphocytes, the cells were isolated from total splenocytes as described above and cultured 5 * 10 5 cells/well at a density of about 7 * 10 5 /cm 2 for 72 h at 37 °C, 5% CO 2 in TexMACS medium (Miltenyi Biotec, 130-097-196) with 10% Dialyzed FBS (Gibco 26400-044), 1% Penicillin-Streptomycin (Gibco, 15140-122), 55 μM β-mercaptoethanol (Gibco, 31350-010) in 48 well pre-coated with CD3ε antibody (pure-functional grade, Miltenyi Biotec, 130-092-973) and then stimulated with CD28 antibody (pure-functional grade, Miltenyi Biotec, 130-093-182).

Techniques: Immunocytochemistry, Expressing, Isolation, Staining, Confocal Microscopy, Western Blot, Phospho-proteomics, Inhibition, Purification, MANN-WHITNEY

Journal: Science Advances

Article Title: B7-H3×4-1BB bispecific antibody augments antitumor immunity by enhancing terminally differentiated CD8 + tumor-infiltrating lymphocytes

doi: 10.1126/sciadv.aax3160

Figure Lengend Snippet: ( A and B ) Dose-dependent costimulatory activity of hIgG1 isotype, 1D8, B5, and B5×1D8 on CD8 T cells stimulated with anti-CD3ε (1 μg/ml) and irradiated MC38 hB7-H3 . Flow cytometric analysis of surface expression on CD8α + T cells (A) and IFN-γ secretion by ELISA (B) 72 hours after stimulation. ( C ) EC 50 values for each parameter. ( D ) Flow cytometric analysis of surface expression on CD8α + T cells stimulated with anti-CD3ε (1 μg/ml) and irradiated wild-type MC38 or MC38 hB7-H3 with indicated antibodies (1 μg/ml) 72 hours after stimulation. ( E ) Representative ex vivo fluorescence images of spleen (S) and tumor (T) (left), and tumor/spleen ratio (right) from MC38 hB7-H3 tumor–bearing mice 24 hours after intravenous injection of 37.5 μg of 680XL-labeled mAb (1D8 and B5) or 50.0 μg of 680XL-labeled B5×1D8 ( n = 3 per group). * P < 0.05; ** P < 0.01; *** P < 0.001; and **** P < 0.0001, two-way analysis of variance (ANOVA) with Bonferroni posttests compared with hIgG1 isotype group (A and B), two-way ANOVA with Bonferroni posttests (D), and one-way ANOVA with Bonferroni’s multiple comparison test (E). ns, not significant.

Article Snippet: Splenocytes (5 × 10 5 ) were placed in a 96-well plate and cultured with In Vivo Ready anti-mouse CD3ε mAb (1 μg/ml) (clone: 145-2C11; Tonbo Biosciences, 40-0031) alone or with purified antibodies at the indicated dose.

Techniques: Activity Assay, Irradiation, Expressing, Enzyme-linked Immunosorbent Assay, Ex Vivo, Fluorescence, Injection, Labeling, Comparison

Journal: Science Advances

Article Title: B7-H3×4-1BB bispecific antibody augments antitumor immunity by enhancing terminally differentiated CD8 + tumor-infiltrating lymphocytes

doi: 10.1126/sciadv.aax3160

Figure Lengend Snippet: ( A ) Dose-dependent costimulatory activity of the urelumab analog and B5×1A10 on Jurkat-NFκB-luc2/h4-1BB reporter cells. Luminescence was measured 6 hours after stimulation. ( B and C ) Dose-dependent costimulatory activity of the urelumab analog, 1A10, and B5×1A10 on PBMCs stimulated with anti-human CD3 (5 μg/ml) and HCC1954 cells. IFN-γ secretion by ELISA (B) and optical cellular density by cell counting kit (C) 72 hours after stimulation. ( D and E ) MC38 hB7-H3 tumor–bearing h4-1BB KI mice ( n = 6 to 8 per group) were treated with indicated antibodies. Black arrows indicate treatment points. Tumor growth curves of individual mice are shown on the right. ( F ) Representative immunohistochemistry images (left) showing CD8 expression (brown) in tumor tissue from mice in (D). Infiltrating CD8 + cell counts in a high-power field (HPF; 20×) at randomly selected tumor areas (right). Nuclear staining with hematoxylin is in blue. Scale bars, 100 μm. * P < 0.05; ** P < 0.01; *** P < 0.001; and **** P < 0.0001, two-way ANOVA with Bonferroni posttests (D and E) and unpaired Student’s t test (F).

Article Snippet: Splenocytes (5 × 10 5 ) were placed in a 96-well plate and cultured with In Vivo Ready anti-mouse CD3ε mAb (1 μg/ml) (clone: 145-2C11; Tonbo Biosciences, 40-0031) alone or with purified antibodies at the indicated dose.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Cell Counting, Immunohistochemistry, Expressing, Staining

Journal: Immunity

Article Title: The E-Id Protein Axis Specifies Innate and Adaptive Lymphoid Cell Fate

doi: 10.1016/j.immuni.2017.04.022

Figure Lengend Snippet: (A) Volcano plots representing RNA-seq reads in sorted ETPs (CD4−CD8−Lin−CD44+CD25−Kit+YFP+) derived from Tcf3fl/flTcf12fl/flIl7rCre fetal thymi. Points represent genes upregulated in Tcf3fl/flTcf12fl/flIl7rCre ETPs > twofold (red; FDR < 0.05 (calculated using raw count values)), genes downregulated in Tcf3fl/flTcf12fl/flIl7rCre ETPs > twofold (blue; P < 0.05) and not significantly changed (grey) as compared to control cells. Selected genes are labelled. (n=3) (B) Expression levels associated with the Notch1, Rag1/Rag2, Ptcra, Cd3d/Cd3e, Tox, Tcf7, and Bcl11b loci, presented as RNA-seq Reads Per Million reads aligned (RPM). (C) Gene Ontology (GO) analysis of significantly elevated gene expression levels in ETPs derived from Tcf3fl/flTcf12fl/flIl7rCre mice using Metascape. Top 15 clusters of GO terms are shown. (D) Heat maps of selected genes are presented to visualize differences in gene expression patterns for control and Tcf3/Tcf12-deficient ETPs isolated from fetal thymi. The heat maps represent different ontology groups, including genes encoding for factors that regulate transcription (left), cytokine/chemokine/surface receptors, signal transduction (middle) as well as cytokines (right). See also Figure S5 and S6.

Article Snippet: Anti-mouse CD3e Biotin , TONBO , 30-0031.

Techniques: RNA Sequencing Assay, Derivative Assay, Expressing, Isolation, Transduction

Journal: Immunity

Article Title: The E-Id Protein Axis Specifies Innate and Adaptive Lymphoid Cell Fate

doi: 10.1016/j.immuni.2017.04.022

Figure Lengend Snippet:

Article Snippet: Anti-mouse CD3e Biotin , TONBO , 30-0031.

Techniques: Recombinant, Staining, Sequencing, Software

Journal: Journal of Extracellular Vesicles

Article Title: High‐Yield Outer Membrane Vesicles Derived From Probiotics as a Nanoplatform for Precise Treatment and Prophylaxis of Pseudomonas aeruginosa Infection

doi: 10.1002/jev2.70194

Figure Lengend Snippet: In vivo immune response and preventive effect activated by FI‐mOMVs. (a) Schematic diagram of immunization, sample collection and challenge schedule. BALB/c mice were immunized s.c. with the indicated formulations on Days 0, 14, 28 ( n = 8), and a PBS control was set up ( n = 5). (b) Serum was collected 7, 14, 21, 28, 35 and 42 days after initial vaccination, and the FI antigen–specific IgG antibody titre was detected by ELISA. (c–e) Detection of antigen‐specific cellular immune response. The spleen lymphocytes were isolated 6 weeks after the initial immunization. The proliferation levels were detected by CCK8 (c), and the expression levels of IFN‐γ (d) and IL‐4 (e) in the culture supernatant were determined by ELISA. (f, g) Percentage of CD3 + CD4 + T lymphocytes (f), and CD3 + CD8 + T lymphocytes (g) in the spleen on Day 42 after immunization ( n = 8). Mice without immunization were used as controls ( n = 5). (h) Survival curves of the mice infected with PAO1 (1 × 10 8 CFU/mouse, i.n.) after different treatments ( n = 12). (i, j) Quantification of the number of bacteria in the lung (i) and the pathological changes of the lung (j) of the mice infected with a sublethal dose of PAO1 (5 × 10 6 CFU/mouse, i.n.) after different treatments ( n = 6). Scale bar, 200 µm. The data are presented as mean ± SEM. Statistical analysis was performed by a two‐tailed unpaired Student's t ‐test (b–g), log‐rank (Mantel–Cox) test (h), or two‐tailed Mann–Whitney U ‐test (i), and differences were considered significant at p < 0.05. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001.

Article Snippet: The cell suspensions (100 μL) were taken and stained with FITC‐conjugated anti‐mouse CD4 (cat# FITC‐65104, Proteintech, USA), PE‐conjugated anti‐mouse CD8 (cat# PE‐65069, Proteintech, USA) and APC‐conjugated anti‐mouse CD3 (cat# APC‐65077, Proteintech, USA) for 30 min at 37°C in the dark.

Techniques: In Vivo, Control, Enzyme-linked Immunosorbent Assay, Isolation, Expressing, Infection, Bacteria, Two Tailed Test, MANN-WHITNEY