Journal: Virology

Article Title: The V domain of dog PVRL4 (nectin-4) mediates canine distemper virus entry and virus cell-to-cell spread.

doi: 10.1016/j.virol.2014.02.014

Figure Lengend Snippet: Fig. 2. Cell surface expression of chimeric PVRL molecules (dog.PVRL4, dog.PVRL4/VhPVRL1 and hPVRL1/Vdog.PVRL4). The flow cytometric histograms depict the cell surface expression profile of PVRL4 and PVRL1 in the corresponding cell line (isotype ctrl antibody, IgG – shaded; PVRL4 antibody, PVRL4 – blue; PVRL1 antibody, PVRL1 – green). Vero stable cell lines were incubated with either a phycoerythrin (PE)-conjugated mouse monoclonal antibody specific for human PVRL4, a PE-conjugated mouse IgG2a control antibody, an allophycocyanin (APC)-conjugated mouse monoclonal antibody specific for human PVRL1 or an APC-conjugated mouse IgG2B control antibody. The Y-axis represents cell counts and the X-axis represents fluorescence intensity (FL8: APC; FL2: PE).

Article Snippet: Briefly, cells were washed with PBS, non-enzymatically dissociated and subsequently blocked in 2.5 μg of normal human IgG (R&D Systems) for 10 min on ice followed by the addition of 10 μl of either PE-conjugated monoclonal antibody against human PVRL4 27-351 (R&D Systems FAB2659P) or PE-conjugated mouse IgG2B isotype control (R&D Systems IC0041P), and APC-conjugated mouse monoclonal antibody against human PVRL1 31-334 (R&D Systems FAB2880A) or APC-conjugated mouse IgG2A isotype control (R&D Systems IC003A) antibodies for 45 min on ice.

Techniques: Expressing, Stable Transfection, Incubation, Control

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