Journal: Journal of Crohn's & Colitis

Article Title: The microRNA Expression in Crypt-Top and Crypt-Bottom Colonic Epithelial Cell Populations Demonstrates Cell-Type Specificity and Correlates with Endoscopic Activity in Ulcerative Colitis

doi: 10.1093/ecco-jcc/jjae108

Figure Lengend Snippet: Sequencing data reveal differentially expressed miRNAs in colonic epithelial cells of patients with active and quiescent UC. [A] FACS of colonic epithelial cells and distribution of crypt-top CD44 − CD66a + and crypt-bottom CD44 + CD66a − epithelial cell types in inflammatory [aUC] [ n = 16] and non-inflammatory [qUC and HC] [ n = 15 and 17] colon tissues. Each dot represents a sample from the patients of the second study group. Mean ± SD of each group is represented by vertical lines. To compare the groups a non-parametric Mann–Whitney U test was performed, * p < 0.05. [B] MDS plot showing the similarity structure of the miRNA transcriptomes in crypt-top [CD66a + ] and crypt-bottom [CD44 + ] colonic epithelial cell populations in active UC [aUC] [ n = 16], quiescent UC [qUC] [ n = 15], and HC [ n = 17] based on normalized expression values. Dots represent samples shaped by cell population. Dot colours represent condition. The centroid of ellipses corresponds to the condition group mean; the shapes are defined by covariance within the group. [C, D] Volcano plots of differentially expressed miRNAs in crypt-top [CD66a + ] and crypt-bottom [CD44 + ] colonic epithelial cell populations in aUC [ n = 16], qUC [ n = 15], and HC [ n = 17]. Colours indicate significantly [FDR < 0.05] differentially expressed miRNAs with an absolute value of log 2 FC > 1 between compared groups. [E–G] Venn diagrams representing the numbers of commonly and uniquely differentially expressed miRNAs in [E] crypt-bottom [CD44 + ] and [F] crypt-top [CD66a + ] epithelial cell populations in different UC activity, and [G] between crypt-bottom and crypt-top cells in the same condition. [H] Overrepresented pathways with the top five FDR values between crypt-top [CD66a + ] and crypt-bottom [CD44 + ] colonic epithelial cell populations during aUC [ n = 16], qUC [ n = 15], and HC [ n = 17] identified by miRNA–target gene set enrichment analysis. Dot size represents the number of miRNA gene–target counts in the significantly enriched [FDR < 0.05] GO categories.

Article Snippet: Antibodies used in this study were selected based on previous study and included: mouse anti-human CD326/EpCAM-FITC [clone VU-1D9, Life Technologies], mouse anti-human CD44-APC [clone G44-26, BD Biosciences], and mouse anti-human CD66a-PE [clone 283340, R&D Systems].

Techniques: Sequencing, MANN-WHITNEY, Expressing, Activity Assay

Journal: Journal of Crohn's & Colitis

Article Title: The microRNA Expression in Crypt-Top and Crypt-Bottom Colonic Epithelial Cell Populations Demonstrates Cell-Type Specificity and Correlates with Endoscopic Activity in Ulcerative Colitis

doi: 10.1093/ecco-jcc/jjae108

Figure Lengend Snippet: WGCNA in colonic epithelial cell populations reveals miRNA co-expression modules which reflect endoscopic activity of UC. [A] Network displaying identified co-expression modules [M1 and M2] in crypt-top [CD66a] and crypt-bottom [CD44] colonic epithelial cells of patients with active [aUC] [ n = 16], quiescent UC [qUC] [ n = 15], and control individuals [ n = 17]. The colour and size of the node represents distinct modules and strength of connectivity, respectively. [B] Dot plot showing normalized enrichment score [NES] of modules M1 and M2 in crypt-top [CD66a] and crypt-bottom [CD44] colonic epithelial cells of patients with active UC, quiescent UC, and control individuals. The colour and size of the dot represents the value and absolute value of NES, respectively. The box marks significant value [ p adj, < 0.05]. Plots [C] and [E] show the correlation between module M1 eigengene value and Mayo endoscopic activity in crypt-bottom [CD44] and crypt-top [CD66a] colonic epithelial cells as well as colon tissue, respectively. rho—Spearman correlation coefficient. Each dot represents a different sample. Plots [D] and [F] show the AUC-ROC curves reflecting the performance of M1 module eigengene value at distinguishing between active UC [aUC] and quiescent UC [qUC] in crypt-bottom [CD44] and crypt-top [CD66a] colonic epithelial cells as well as in colon tissue, respectively.

Article Snippet: Antibodies used in this study were selected based on previous study and included: mouse anti-human CD326/EpCAM-FITC [clone VU-1D9, Life Technologies], mouse anti-human CD44-APC [clone G44-26, BD Biosciences], and mouse anti-human CD66a-PE [clone 283340, R&D Systems].

Techniques: Expressing, Activity Assay, Control

Journal: Gastroenterology

Article Title: T Cell CEACAM1-TIM-3 Crosstalk Alleviates Liver Transplant Injury in Mice and Humans.

doi: 10.1053/j.gastro.2023.07.004

Figure Lengend Snippet: Figure 1. Recipient CC1 signaling mitigates hepatic IRI and suppresses NF-kB p65 in mouse OLT. Mouse WT livers subjected to 18 hours of cold storage were transplanted into WT or CC1KO syngeneic recipients (n ¼ 6–8/group). OLT/serum samples were analyzed 6 hours after reperfusion. The sham group (n ¼ 5) underwent the same procedures except for OLT. (A) Representative H&E and TUNEL staining. DAPI, 40,6-diamidino-2-phenylindole. Scale bars ¼ 100 mm. (B) Suzuki’s histologic grading of liver IRI and quantification of TUNELþ cells/HPF. (C) sAST/sALT and (D) lactate dehydrogenase (LDH) levels (U/L). Quantitative reverse transcription polymerase chain reaction–assisted OLT detection of (E) IFN-gamma, IL6, IL17, and gran- zyme B/perforin 1, and (F) TLR4, IL1b, and TNF-a (n ¼ 6–7/group). Data normalized to hypoxanthine guanine phosphor- ibosyltransferase gene expression. (G) Western blot–assisted detection of phosphorylated (p)-IkBa, IkBa, p-NF-kB p65, NF-kB p65, and vinculin (VCL). The relative intensity ratio of p-IkBa/IkBa and p-NF-kB p65/NF-kB p65 (n ¼ 4/group) is shown. Data are shown as mean ± standard error of the mean *P < .05, **P < .01, Student t test.

Article Snippet: Human liver samples were stained with rabbit anti-CD4 (ab133616/ EPR6855, Abcam) and rat anti-CC1 antibody (MAB6480/ 723629, R&D Systems).

Techniques: TUNEL Assay, Staining, Reverse Transcription, Polymerase Chain Reaction, Gene Expression, Western Blot

Journal: Gastroenterology

Article Title: T Cell CEACAM1-TIM-3 Crosstalk Alleviates Liver Transplant Injury in Mice and Humans.

doi: 10.1053/j.gastro.2023.07.004

Figure Lengend Snippet: Figure 2. CC1 signaling enhances TIM-3 expression and suppresses CD4þ T-cell inflammatory signature. (A) Representative (n ¼ 4/group) flow cytometry of CC1 and TIM-3 expression in peripheral blood lymphocytes from naive WT and CC1KO mice. TIM-3þ frequency in (B) WT CC1þ and CC1 CD4þ T cells and in (C) WT and CC1KO CD4þ and CD8þ T cells. (D) Repre- sentative (n ¼ 4/group) flow cytometry of CC1 and TIM-3 expression in activated peripheral blood lymphocytes from WT and CC1KO mice subjected to hepatic IRI. TIM-3þ frequency in (E) WT CC1þ and CC1 CD4þ T cells and in (F) WT and CC1KO CD4þ and CD8þ T cells. (G) CC1þ frequency in naive vs post-IRI in WT CD4þ T cells. (H) TIM-3þ frequency in naive vs post-IRI in WT and CC1KO CD4þ T cells. (I) Quantitative reverse-transcription polymerase chain reaction–assisted detection of IFN-g, T-box protein (T-bet) expressed in T cells, IL6, IL17, IL22, and TNF-a in CD4þ T cells from WT and CC1KO mice before and after stimulation (n ¼ 6/group). White square: WT; black square: CC1KO mouse. Data shown as mean ± standard error of the mean. *P < .05, ****P < .0001, Student t test.

Article Snippet: Human liver samples were stained with rabbit anti-CD4 (ab133616/ EPR6855, Abcam) and rat anti-CC1 antibody (MAB6480/ 723629, R&D Systems).

Techniques: Expressing, Cytometry, Reverse Transcription, Polymerase Chain Reaction

Journal: Gastroenterology

Article Title: T Cell CEACAM1-TIM-3 Crosstalk Alleviates Liver Transplant Injury in Mice and Humans.

doi: 10.1053/j.gastro.2023.07.004

Figure Lengend Snippet: Figure 4. Enhanced T cell–specific TIM-3 alleviates IRI-OLT and suppresses Kupffer cell NF-kB p65 in CC1-deficient re- cipients. WT livers after 18 hours of cold storage were transplanted into WT, CC1KO, T cell–specific TIM-3Tg/CC1KO, and TIM-3Tg/CC1KO þ anti–TIM-3 antibody (n ¼ 6–8/group). OLT/serum samples were analyzed at 6 hours. (A) Representative H&E and TUNEL staining. DAPI, 40,6-diamidino-2-phenylindole. Scale bars ¼ 100 mm. (B) Suzuki’s histologic grading of liver IRI and quantification of TUNELþ cells/HPF. (C) sAST/sALT (U/L). Quantitative reverse-transcription polymerase chain reaction–assisted detection of (D) IFN-gamma, IL6, and IL17, and (E) TLR4, IL-1b, and TNF-a in OLT (n ¼ 6/group). Data in D and E were normalized to hypoxanthine guanine phosphoribosyltransferase gene expression. (F) Western blot-assisted detection of CC1, phosphorylated (p)-NF-kB p65, NF-kB p65, and vinculin (VCL). The relative intensity ratio of p-NF-kB p65/NF-kB p65 (n ¼ 3–4/group) is shown. (G) Representative NF-kB staining in OLT. Arrows indicate nuclear NF-kB locali- zation in nonparenchymal cells. Scale bars ¼ 100 mm. (H) Representative C-type lectin domain family 4 member F (CLEC4F; Kupffer cell) and p-NF-kB p65 staining. Arrowheads indicate Kupffer cells augmenting p-NF-kB p65. Scale bars ¼ 100 mm (left panels) and 20 mm (enlarged images). Data are shown as mean ±standard error of the mean. *P < .05, **P < .01, ***P < .01, Student t test.

Article Snippet: Human liver samples were stained with rabbit anti-CD4 (ab133616/ EPR6855, Abcam) and rat anti-CC1 antibody (MAB6480/ 723629, R&D Systems).

Techniques: TUNEL Assay, Staining, Reverse Transcription, Polymerase Chain Reaction, Gene Expression, Western Blot

Journal: Gastroenterology

Article Title: T Cell CEACAM1-TIM-3 Crosstalk Alleviates Liver Transplant Injury in Mice and Humans.

doi: 10.1053/j.gastro.2023.07.004

Figure Lengend Snippet: Figure 5. Donor liver CC1 deficiency compromises T cell–specific TIM-3 regulation in CC1-deficient recipients. (A) CC1KO livers after 18 hours of cold storage were transplanted into CC1KO or TIM-3Tg/CC1KO mice. OLT/serum samples were analyzed at 6 hours (n ¼ 6/group). The sham group (n ¼ 5) underwent the same procedures, except for OLT. (B) Representative H&E staining. Scale bars ¼ 100 mm. (C) Suzuki’s histologic grading of liver IRI and sAST/sALT (U/L). (D) Representative C-type lectin domain family 4 member F (CLEC4F; Kupffer cells) and phosphorylated (p)-NF-kB p65 staining in OLT. DAPI, 40,6- diamidino-2-phenylindole. Arrowheads indicate p-NF-kB p65þ cells. Scale bars ¼ 100 mm. Quantative reverse-transcription polymerase chain reaction–assisted detection of (E) IFN-gamma, IL6, and IL17 (n ¼ 6/group), and (F) TLR4, IL-1b, and TNF-a (n ¼ 6/group) in OLT. Data were normalized to hypoxanthine guanine phosphoribosyltransferase (HPRT) gene expression. (G) Western blot–assisted detection of CC1, p-NF-kB p65, NF-kB p65, and b-actin. The relative intensity ratio of p- NF-kB p65/NF-kB p65 (n ¼ 3/group) is shown. Data are shown as mean ± standard error of the mean. ***P < .001, Student t test.

Article Snippet: Human liver samples were stained with rabbit anti-CD4 (ab133616/ EPR6855, Abcam) and rat anti-CC1 antibody (MAB6480/ 723629, R&D Systems).

Techniques: Staining, Reverse Transcription, Polymerase Chain Reaction, Gene Expression, Western Blot

Journal: Gastroenterology

Article Title: T Cell CEACAM1-TIM-3 Crosstalk Alleviates Liver Transplant Injury in Mice and Humans.

doi: 10.1053/j.gastro.2023.07.004

Figure Lengend Snippet: Figure 6. Perioperative increase of CC1 promotes anti-inflammatory phenotype in human OLT. (A) Pretransplant (after cold storage) and posttransplant (2 hours after reperfusion) hepatic biopsy specimens were collected from OLT patients. Post-/pre- CC1 ratios were analyzed at the gene (n ¼ 27) and protein (n ¼ 50) levels. Relationship between post-/pre-CC1 gene ratio and (B) CD154, CD28, IFN-gamma, and IL-17, (C) TLR2, TLR4, TLR9, and CD68, and (D) cathepsin G and HO-1 gene expression with b-actin normalization (n ¼ 27), *P < .05, **P < .01; nonparametric Spearman’s method.

Article Snippet: Human liver samples were stained with rabbit anti-CD4 (ab133616/ EPR6855, Abcam) and rat anti-CC1 antibody (MAB6480/ 723629, R&D Systems).

Techniques: Gene Expression

Journal: Gastroenterology

Article Title: T Cell CEACAM1-TIM-3 Crosstalk Alleviates Liver Transplant Injury in Mice and Humans.

doi: 10.1053/j.gastro.2023.07.004

Figure Lengend Snippet: Figure 7. Perioperative increase of CC1 attenuates hepatocellular injury and improves rejection-free human OLT survival. Post-/pre- CC1 ratios were assessed by Western blots with b-actin normalization. (A) OLT patients were divided into low (n ¼ 25) and high (n ¼ 25) post-/pre-CC1 ratio groups, based on the median value of the CC1 ratio (cutoff ¼ 1.05). (B) Representative Western blots and case patient-related clinical parameters (case patients 1 and 2: low post-/pre-CC1 ratio, case patients 3 and 4: high post-/pre-CC1 ratio). (C) sAST/sALT at POD 17. (D) Representative CD4/CC1 staining in OLT. Arrows: CC1-negative CD4þ T cells; arrowheads: CC1-positive CD4þ T cells. Scale bars ¼ 100 mm. DAPI, 40,6-diamidino-2-phenylindole. (E) Incidence of EAD. (F) The cumulative rejection rate (Kaplan-Meier method). The solid line indicates high and dotted line low post-/pre-CC1 ratio in human OLT. Data shown as mean ± standard error of the mean. *P < .05, Mann-Whitney U test in C, Fisher’s exact test in E, and log-rank test in F.

Article Snippet: Human liver samples were stained with rabbit anti-CD4 (ab133616/ EPR6855, Abcam) and rat anti-CC1 antibody (MAB6480/ 723629, R&D Systems).

Techniques: Western Blot, Staining, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Neutrophils Infiltration in the Tongue Squamous Cell Carcinoma and Its Correlation with CEACAM1 Expression on Tumor Cells

doi: 10.1371/journal.pone.0089991

Figure Lengend Snippet: The expression of CEACAM1 on different TSCC groups and its relationship with clinicopathologic features.

Article Snippet: The membrane was blocked with 5% non-fat milk for 1 h, and incubated with monoclonal mouse anti-human CEACAM1 (dilution 1∶500, RD; MAB2244, USA) antibodies for 2 h at room temperature, followed by washing with TBST (TBS containing 0.1% Tween 20) for 5 min five times.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Neutrophils Infiltration in the Tongue Squamous Cell Carcinoma and Its Correlation with CEACAM1 Expression on Tumor Cells

doi: 10.1371/journal.pone.0089991

Figure Lengend Snippet: The relationship of CEACAM1 expression and density of neutrophils.

Article Snippet: The membrane was blocked with 5% non-fat milk for 1 h, and incubated with monoclonal mouse anti-human CEACAM1 (dilution 1∶500, RD; MAB2244, USA) antibodies for 2 h at room temperature, followed by washing with TBST (TBS containing 0.1% Tween 20) for 5 min five times.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Neutrophils Infiltration in the Tongue Squamous Cell Carcinoma and Its Correlation with CEACAM1 Expression on Tumor Cells

doi: 10.1371/journal.pone.0089991

Figure Lengend Snippet: Univariate and Multivariate survival analysis in TSCC patients.

Article Snippet: The membrane was blocked with 5% non-fat milk for 1 h, and incubated with monoclonal mouse anti-human CEACAM1 (dilution 1∶500, RD; MAB2244, USA) antibodies for 2 h at room temperature, followed by washing with TBST (TBS containing 0.1% Tween 20) for 5 min five times.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Neutrophils Infiltration in the Tongue Squamous Cell Carcinoma and Its Correlation with CEACAM1 Expression on Tumor Cells

doi: 10.1371/journal.pone.0089991

Figure Lengend Snippet: Primer sequences and size of PCR products.

Article Snippet: The membrane was blocked with 5% non-fat milk for 1 h, and incubated with monoclonal mouse anti-human CEACAM1 (dilution 1∶500, RD; MAB2244, USA) antibodies for 2 h at room temperature, followed by washing with TBST (TBS containing 0.1% Tween 20) for 5 min five times.

Techniques:

Journal: PLoS ONE

Article Title: Neutrophils Infiltration in the Tongue Squamous Cell Carcinoma and Its Correlation with CEACAM1 Expression on Tumor Cells

doi: 10.1371/journal.pone.0089991

Figure Lengend Snippet: A: There were more CD15+ neutrophils (black arrow) in TSCC tissues (b, c, d) than in peritumoral tissues (a). In tumors areas, some neutrophils lied in the stroma of tumors (b), some were located within the carcinoma nests (c), and some infiltrated in the borderline of tumor invasion (d). (a: 100×; b, c, d: 400×). B: The expression of CEACAM1 in peritumoral tissues was negative or weak (a). In TSCC tissues, CEACAM1 expression was obviously stronger than that in peritumoral tissues and located mainly in the cytoplasm of tumor cells (b). (a: 100×; b: 400×). C: The relationship of CEACAM1 expression on tumor cells and infiltration of neutrophils. In strong CEACAM1 expression tumors, there were more neutrophils (black arrow) (a); While in negative or weak CEACAM1 expression tumors, there were relatively fewer neutrophils (b). (a, b: 200×, for CEACAM1).

Article Snippet: The membrane was blocked with 5% non-fat milk for 1 h, and incubated with monoclonal mouse anti-human CEACAM1 (dilution 1∶500, RD; MAB2244, USA) antibodies for 2 h at room temperature, followed by washing with TBST (TBS containing 0.1% Tween 20) for 5 min five times.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Neutrophils Infiltration in the Tongue Squamous Cell Carcinoma and Its Correlation with CEACAM1 Expression on Tumor Cells

doi: 10.1371/journal.pone.0089991

Figure Lengend Snippet: A: The relationship between cumulative cancer-related survival of patients with resectable TSCC and density of neutrophils in tumor tissues. Patients with high density of neutrophils in tumors had significantly reduced cancer-related survival compared to low neutrophils density group ( P = 0.020, Log-rank test). B: The relationship between cumulative cancer-related survival of patients with resectable TSCC and CEACAM1 expression on tumor cells. Patients with moderate or strong expression of CEACAM1 had significantly reduced cancer-related survival compared to negative or weak CEACAM1 expression group ( P = 0.032, Log-rank test).

Article Snippet: The membrane was blocked with 5% non-fat milk for 1 h, and incubated with monoclonal mouse anti-human CEACAM1 (dilution 1∶500, RD; MAB2244, USA) antibodies for 2 h at room temperature, followed by washing with TBST (TBS containing 0.1% Tween 20) for 5 min five times.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Neutrophils Infiltration in the Tongue Squamous Cell Carcinoma and Its Correlation with CEACAM1 Expression on Tumor Cells

doi: 10.1371/journal.pone.0089991

Figure Lengend Snippet: A: Fluorescence microscopic observation of Lv transfection efficientcy. a: CC1-4L-Lv group; b: CC1-4S-Lv group; c: Vector-Lv group. (a, b, c 200×). B: qRT- PCR results of Lv transfection. The results showed that CEACAM1-4L mRNA expression was obviously higher in CC1-4L-Lv transfection group than the other three groups (a), and CEACAM1-4S mRNA expression was the same as 4L (b). C: Western blot results for Lv transfection. a: β-actin; b: CEACAM1-4L (the above band) and -4S (the lower band). Results showed that CEACAM1-4L protein expression was obviously stronger in CC1-4L-Lv transfection group than the other three groups. CEACAM1-4S protein expression was the same as 4L. (Note: CEACAM1-4L protein was slightly bigger than CEACAM1-4S).

Article Snippet: The membrane was blocked with 5% non-fat milk for 1 h, and incubated with monoclonal mouse anti-human CEACAM1 (dilution 1∶500, RD; MAB2244, USA) antibodies for 2 h at room temperature, followed by washing with TBST (TBS containing 0.1% Tween 20) for 5 min five times.

Techniques: Fluorescence, Transfection, Plasmid Preparation, Quantitative RT-PCR, Expressing, Western Blot

Journal: Cell reports

Article Title: SOX10 requirement for melanoma tumor growth is due, in part, to immune-mediated effects

doi: 10.1016/j.celrep.2021.110085

Figure Lengend Snippet: (A) Parental mouse melanoma YUMM1.1 cells and Sox10 knockout clones were treated with or without 100 ng/mL IFN γ , and HVEM expression was probed by flow cytometry staining. Mean fluorescence intensity (MFI) was quantified. (B) Parental human melanoma MeWo cells and SOX10 knockout clones were treated with or without 100 ng/mL IFN γ . HVEM expression was probed by flow cytometry staining, and MFI was quantified. (C) Parental mouse melanoma YUMM1.1 cells and Sox10 knockout clones were treated with or without 100 ng/mL IFN γ . CEACAM1 expression was probed by flow cytometry staining, and MFI was quantified. (D) Parental human melanoma MeWo cells and SOX10 knockout clones were treated with or without 100 ng/mL IFN γ . CEACAM1 expression was probed by flow cytometry staining, and MFI was quantified. (E) CEACAM1 expression was probed by western blot in parental mouse melanoma YUMM1.1 cells and Sox10 knockout clones. The arrow indicates a nonspecific band ("n.s."). (F) CEACAM1 expression was probed by western blot in parental human melanoma MeWo cells and SOX10 knockout clones. All data in this figure represent 3 biological replicates; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Mouse CEACAM1 polyclonal antibody (#AF6480) was purchased from R&D Systems.

Techniques: Knock-Out, Clone Assay, Expressing, Flow Cytometry, Staining, Fluorescence, Western Blot

Journal: Cell reports

Article Title: SOX10 requirement for melanoma tumor growth is due, in part, to immune-mediated effects

doi: 10.1016/j.celrep.2021.110085

Figure Lengend Snippet: (A) RNA-seq from the melanoma TCGA dataset was visualized using cBioportal, and the mRNA expression level of HVEM ( TNFRSF14 ) was plotted versus SOX10 expression ( ; ). Shown are values batch normalized from Illumina HiSeq_RNASeqV2. SOX10 mutation type, structural variant, and copy number are indicated for each sample. (B) RNA-seq from the melanoma TCGA dataset was visualized using cBioportal, and the mRNA expression level of CEACAM1 was plotted versus SOX10 express ion ( ; ). Shown are values batch normalized from Illumina HiSeq_RNASeqV2. SOX10 mutation type, structural variant, and copy number are indicated for each sample. (C) Single-cell RNA-seq data of malignant cells were visualized by t-distributed stochastic neighbor embedding (tSNE) plots by using the Single Cell portal ( https://singlecell.broadinstitute.org/single_cell ), whereby each cluster represents a distinct melanoma tumor and each circle represents an individual cell.

Article Snippet: Mouse CEACAM1 polyclonal antibody (#AF6480) was purchased from R&D Systems.

Techniques: RNA Sequencing, Expressing, Mutagenesis, Variant Assay

Journal: Cell reports

Article Title: SOX10 requirement for melanoma tumor growth is due, in part, to immune-mediated effects

doi: 10.1016/j.celrep.2021.110085

Figure Lengend Snippet: (A) HVEM was expressed in YUMM1.1 parental and CR. #1.41 Sox10 knockout cells by lentiviral transduction. HVEM expression was validated by flow cytometry staining. Data are representative of 3 biological replicates. An unstained control is shown in light gray. (B) YUMM1.1 parental, CR. #1.41 Sox10 knockout cells, or HVEM-overexpressing cells were injected into BL6 mice, and tumors were measured by digital caliper every 2–3 days. Data were collected from 7 mice per group. (C) Related to (B), time-to-tumor onset was tracked. Tumors were considered fully formed when they reached ~50 mm 3 . (D) Two individual clones were generated from CRISPR-Cas9 knockout of Ceacam1 in YUMM1.1 cells. CEACAM1 surface expression was evaluated by flow cytometry. Data are representative of 3 biological replicates. An unstained control is shown in light gray. (E) YUMM1.1 parental, CR. #1.8, or CR. #1.30 Ceacam1 knockout cells were injected into BL6 mice, and tumors were measured by caliper every 2–3 days. Data were collected from 10 mice per group. (F) Related to (E), time-to-tumor onset was tracked. Tumors were considered fully formed when they reached ~50 mm 3 .**p < 0.01, ***p < 0.001.

Article Snippet: Mouse CEACAM1 polyclonal antibody (#AF6480) was purchased from R&D Systems.

Techniques: Knock-Out, Transduction, Expressing, Flow Cytometry, Staining, Control, Injection, Clone Assay, Generated, CRISPR

Journal: Cell reports

Article Title: SOX10 requirement for melanoma tumor growth is due, in part, to immune-mediated effects

doi: 10.1016/j.celrep.2021.110085

Figure Lengend Snippet:

Article Snippet: Mouse CEACAM1 polyclonal antibody (#AF6480) was purchased from R&D Systems.

Techniques: Control, Recombinant, CRISPR, Knockdown, Western Blot, Generated, Software