mouse ceacam1 polyclonal antibody  (R&D Systems)

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

Journal: Metabolism: clinical and experimental

Article Title: Hyperinsulinemia drives hepatic insulin resistance in male mice with liver-specific Ceacam1 deletion independently of lipolysis.

doi: 10.1016/j.metabol.2019.01.008

Figure Lengend Snippet: Fig. 1. Tissue-specific expression of mouse CEACAM1 (mCC1). (A) Primary hepatocytes from Alb–Cc1+/+ (white bar), Alb+Cc1+/+ (light grey bar), Alb–Cc1fl/fl(dark grey bar) and Alb+Cc1fl/fl(black bar) were isolated and analyzed by qRT-PCR in triplicate to assess mouse Ceacam1 mRNA level. Values are expressed as mean ± SEM; *P b 0.05 vs Alb– Cc1+/+, Alb+Cc1+/+ and Alb–Cc1fl/fl. (B) Western blot analysis of mouse (mCC1) CEACAM1 protein content in (i) liver and kidney, and (ii) hypothalamus and heart was performed by immunoblotting (Ib) the upper portion of the blot with a polyclonal antibody against CEACAM1 (α-CC1) and the lower portion with α-Tubulin to normalize for protein loading. Gels represent more than two experiments (different mice/genotype).

Article Snippet: Cells were lysed in 1%Triton-X and proteins immunoprecipitated with streptavidin (Fisher Scientific, Waltham, MA), and immunoblotted with 1:1000 of insulin receptor alpha (IRα) antibody (N-20, Santa Cruz) or a custom-made mouse Ab3759 polyclonal antibody against CEACAM1 [20], followed by horseradish peroxidase-conjugated mouse anti-rabbit IgG antibody (Jackson Immunoresearch, West Grove, PA) and enhanced chemiluminescence (ECL, Amersham Pharmacia, Sunnyvale, CA).

Techniques: Expressing, Isolation, Quantitative RT-PCR, Western Blot

Journal: Metabolism: clinical and experimental

Article Title: Hyperinsulinemia drives hepatic insulin resistance in male mice with liver-specific Ceacam1 deletion independently of lipolysis.

doi: 10.1016/j.metabol.2019.01.008

Figure Lengend Snippet: Fig. 8. Hyperphagia and systemic insulin resistance. (A) Pair-feeding experiments were performed on 7–8 month-old mice with some (i) Alb+Cc1fl/flbeing fed ad libitum (AL) and others being subjected to caloric restriction (CR) for 2 weeks to decrease their body mass to the level of Ad libitum fed controls (n = 6/genotype/feeding group). (ii) At the end of the feeding period insulin tolerance was determined. (AL)Alb–Cc1+/+ (white circle), (AL)Alb+Cc1+/+ (light grey triangle), (AL)Alb–Cc1fl/fl(dark grey square), (AL)Alb +Cc1fl/fl(black circle) and (CR)Alb+Cc1fl/fl(hatched circle). Values are expressed as mean ± SEM at each time point. *P b 0.05 vs (AL)Alb–Cc1+/+, (AL)Alb+Cc1+/+, (AL) Alb–Cc1fl/fland (CR)Alb+Cc1fl/fl. (B) Ceacam1 (Cc1) mRNA content was analyzed by qRT- PCR in triplicate in liver (i) and hypothalamus (ii) of Alb–Cc1fl/fl(grey bar) and Alb +Cc1fl/fl(black bar) mice aged 2–9 months (n = 5/genotype/age group). (iii) Cc2 mRNA was analyzed in the hypothalamus of Alb–Cc1fl/fl(grey bar) and Alb+Cc1fl/fl(black bar) mice aged 2–9 months, as in (ii). Values are expressed as mean ± SEM. *P b 0.05 vs Alb– Cc1fl/flof the same age group. †P ≤0.05 vs mice at the earliest age examined.

Article Snippet: Cells were lysed in 1%Triton-X and proteins immunoprecipitated with streptavidin (Fisher Scientific, Waltham, MA), and immunoblotted with 1:1000 of insulin receptor alpha (IRα) antibody (N-20, Santa Cruz) or a custom-made mouse Ab3759 polyclonal antibody against CEACAM1 [20], followed by horseradish peroxidase-conjugated mouse anti-rabbit IgG antibody (Jackson Immunoresearch, West Grove, PA) and enhanced chemiluminescence (ECL, Amersham Pharmacia, Sunnyvale, CA).

Techniques: Quantitative RT-PCR