Journal: Immunobiology

Article Title: c-Rel is dispensable for the differentiation and functional maturation of M cells in the follicle-associated epithelium

doi: 10.1016/j.imbio.2016.09.008

Figure Lengend Snippet: Primers used for RT-qPCR analysis.

Article Snippet: For the detection of SpiB in paraformaldehyde-fixed sections, antigen retrieval was performed with citrate buffer (pH 7.0, 121 °C, 5 min) prior to immunostaining with sheep anti-mouse SpiB polyclonal antibody (R&D Systems).

Techniques:

Journal: Immunobiology

Article Title: c-Rel is dispensable for the differentiation and functional maturation of M cells in the follicle-associated epithelium

doi: 10.1016/j.imbio.2016.09.008

Figure Lengend Snippet: SpiB expression in the FAE is unaffected in the absence of c-Rel. (A) IHC analysis of SpiB expression (green) in the FAE of c-Rel −/− and wild-type (WT) control mice. Boxed areas in upper panels are shown at higher magnification in the lower panels. Broken lines indicate the FAE boundaries. Arrows, Spi-B + cell nuclei in the FAE. (B) Morphometric analysis showed that the number of SpiB + cells in the FAE of c-Rel −/− and WT were similar. (C) RT-qPCR analysis suggested there was no significant difference in the expression of Spib mRNA levels in Peyer’s patches from c-Rel −/− or WT mice. Gene expression data are normalised so that the mean level in samples from WT mice was 1.0. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: For the detection of SpiB in paraformaldehyde-fixed sections, antigen retrieval was performed with citrate buffer (pH 7.0, 121 °C, 5 min) prior to immunostaining with sheep anti-mouse SpiB polyclonal antibody (R&D Systems).

Techniques: Expressing, Control, Quantitative RT-PCR, Gene Expression

Journal: Molecular Cancer Research

Article Title: Norepinephrine Promotes the β1-Integrin–Mediated Adhesion of MDA-MB-231 Cells to Vascular Endothelium by the Induction of a GROα Release

doi: 10.1158/1541-7786.mcr-11-0130

Figure Lengend Snippet: Figure 6. Expression and function of the chemokine receptors CXCR1 and CXCR2 in tumor cell adhesion to HMVECs under flow conditions. A, the expression of the chemokine receptors CXCR1 and CXCR2 was investigated by PCR. b-Actin served as loading control. The standard is a 100-bp ladder. B, confirmatory experiments were carried out with PC-3 human prostate carcinoma cells. Left, adhesion of the cells to HMVECs under flow conditions; right, PCR of CXCR1 and CXCR2 expression. C and D, adhesion of MDA-MB-231 cells to HMVECs under flow conditions. C, the cells were pretreated with antibodies blocking the chemokine receptors (aCXCR1/2) or an isotypic control antibody (IgG2a). D, the endothelium was overlaid with GROa and IL-8 at a concentration of 1 mg/mL for 3 minutes before the experiment. Denaturated chemokines were used as control. Graphs in (C) and (D) show mean values and SDs of 3 independent experiments. , statistically significant changes (P < 0.05). Nor, norepinephrine.

Article Snippet: For blocking experiments, MDA-MB231 were preincubated for 10 minutes with monoclonal mouse anti-CXCR1 and anti-CXCR2 (each 5 mg/mL; R&D Systems), mouse IgG2a (10 mg/mL; R&D Systems), mouse anti-b1-integrin clone 4B4 (5 mg/mL; Beckman Coulter), or mouse IgG1 (5 mg/mL; Beckman Coulter).

Techniques: Expressing, Control, Blocking Assay, Concentration Assay

Journal: Molecular Cancer Research

Article Title: Norepinephrine Promotes the β1-Integrin–Mediated Adhesion of MDA-MB-231 Cells to Vascular Endothelium by the Induction of a GROα Release

doi: 10.1158/1541-7786.mcr-11-0130

Figure Lengend Snippet: Figure 7. Inhibition of b1-integrins and NF-kB abrogates the effect of norepinephrine on MDA-MB-231 cell adhesion. A, MDA-MB-231 cells were pretreated with either the b1-integrin–blocking antibody 4B4 or an isotypic control antibody (IgG1). B, HMVECs were preincubated with an NF-kB activation inhibitor (NF-kB Inh); norepinephrine (Nor) was used at 10 mmol/L. Both graphs show mean values and SDs of 3 independent experiments. , statistically significant changes (P < 0.05).

Article Snippet: For blocking experiments, MDA-MB231 were preincubated for 10 minutes with monoclonal mouse anti-CXCR1 and anti-CXCR2 (each 5 mg/mL; R&D Systems), mouse IgG2a (10 mg/mL; R&D Systems), mouse anti-b1-integrin clone 4B4 (5 mg/mL; Beckman Coulter), or mouse IgG1 (5 mg/mL; Beckman Coulter).

Techniques: Inhibition, Blocking Assay, Control, Activation Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: SAMSN1 Is a Tumor Suppressor Gene in Multiple Myeloma 1 2

doi: 10.1016/j.neo.2014.07.002

Figure Lengend Snippet: SAMSN1 expression is reduced in CD138 + PCs of patients with MM and HMCL. (A) SAMSN1 expression (as determined by real-time PCR) is significantly reduced in the BMs of patients with MM ( n = 34) compared with patients with MGUS ( n = 9) and healthy age-matched controls ( n = 5; * P < .05, ** P < .001, one-way ANOVA with Tukey’s multiple comparison test). (B) SAMSN1 expression in CD138 + MACS isolated PCs from patients with MM negatively correlates with BM PC burden ( n = 10, r 2 = 0.6147, P = .0043). (C) In silico analysis of published microarray data. CD138 + PCs were isolated by MACS from 414 patients with MM, 44 patients with MGUS, and 22 age-matched controls. RNA was extracted and analyzed using the Affymetrix U133Plus2.0 microarray platform (GEO Accession Nos GSE4581 and GSE5900). Expression of SAMSN1 is significantly reduced in PCs of patients with MM compared to those of patients with MGUS and normal controls. P < 0.0001, one-way ANOVA with Tukey’s multiple comparison test. (D) Total RNA was extracted from six HMCLs and reverse transcribed. The levels of SAMSN1 expression were assessed by real-time PCR.

Article Snippet: PCs were isolated from flushed long bones and identified using rat anti-mouse CD138 (R&D Systems, Minneapolis, MN) followed by PE-conjugated goat anti-rat IgG.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Comparison, Isolation, In Silico, Microarray, Reverse Transcription

Journal: Chinese Medical Journal

Article Title: Neutralization of Interleukin-9 Decreasing Mast Cells Infiltration in Experimental Autoimmune Encephalomyelitis

doi: 10.4103/0366-6999.204110

Figure Lengend Snippet: Anti-IL-9 antibody treatment decreased mast cell infiltration of the CNS. Purified cells (mainly composed of mast cells) were harvested from the CNS on days 0, and 5, 15, and 20 after EAE immunization. CD45 + CD117 + mast cells were detected by flow cytometry. CNS mast cells maintained a high level throughout the disease course in EAE group. However, mast cells were significantly reduced from day 5, and remained low in anti-IL-9 Abs group, compared with IgG group. Data are shown as mean ± SE. * P < 0.01, anti-IL-9 Abs group versus IgG group. CNS: Central nervous system; EAE: Experimental autoimmune encephalomyelitis; SE: Standard error; IL: Interleukin.

Article Snippet: The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).

Techniques: Purification, Flow Cytometry

Journal: Chinese Medical Journal

Article Title: Neutralization of Interleukin-9 Decreasing Mast Cells Infiltration in Experimental Autoimmune Encephalomyelitis

doi: 10.4103/0366-6999.204110

Figure Lengend Snippet: IL-9 blockade reduced production of chemokine recruiting mast cells in the CNS. Five days after immunization, mRNA expressions of Pecam1, SCF, Vcam-1, CCL2, and CCL5 in CNS tissue of EAE mice were detected by RT-PCR. After IL-9 neutralization, mRNA expressions of CCL5 and Vcam-1 were significantly decreased in anti-IL-9 Abs group, compared with IgG group. * P < 0.01. Pecam1: Platelet and endothelial cell adhesion molecule 1; SCF: Supercoiling factor; Vcam-1: Vascular cell adhesion molecule 1; CCL2: C-C motif chemokine ligand 2; CCL5: C-C motif chemokine ligand 5; CNS: Central nervous system; EAE: Experimental autoimmune encephalomyelitis; IL: Interleukin; RT-PCR: Reverse transcription-polymerase chain reaction; mRNA: Messenger RNA.

Article Snippet: The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Neutralization

Journal: Chinese Medical Journal

Article Title: Neutralization of Interleukin-9 Decreasing Mast Cells Infiltration in Experimental Autoimmune Encephalomyelitis

doi: 10.4103/0366-6999.204110

Figure Lengend Snippet: In vitro , the effect of anti-IL-9 antibody on splenic mast cells. Splenocytes were harvested from experimental autoimmune encephalomyelitis mice 5 days after MOG immunization. After co-culture with anti-IL-9 antibody or anti-mouse IgG for 7 h, mast cell number was counted by flow cytometry. Splenic mast cells cultured with anti-IL-9 antibody showed significantly lower levels in a dose-dependent manner. This trend was particularly evident with anti-IL-9 antibody concentrations up to 20 μg/ml. * P < 0.05; † P < 0.01. IL: Interleukin; MOG: Myelin oligodendrocyte glycoprotein.

Article Snippet: The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).

Techniques: In Vitro, Co-Culture Assay, Flow Cytometry, Cell Culture