Journal: Circulation Research

Article Title: Immune Cell Toll-Like Receptor 4 Is Required for Cardiac Myocyte Impairment During Endotoxemia

doi: 10.1161/01.res.0000144175.70140.8c

Figure Lengend Snippet: Figure 7. Detection of TLR4 protein on wild-type cardiac ven- tricular myocytes (A) and endothelium (B) were isolated from C57BL/6 mice and fixed for FACS analysis. Mean fluorescence measurements for IgG2a (isotype control) and TLR4 on myocytes/endothelium.

Article Snippet: IgG2a-PE was used as an isotype control to TLR4-PE (MTS510, Santa Cruz).

Techniques: Isolation, Control

Journal: Acta Neuropathologica Communications

Article Title: Brain injury drives optic glioma formation through neuron-glia signaling

doi: 10.1186/s40478-024-01735-w

Figure Lengend Snippet: a Following optic nerve crush (ON-CR) at 6 weeks of age, Nf1 flox/flox ; hGFAP-Cre mice at 12 weeks of age have increased numbers of CD3 + cells in their optic nerves relative to those receiving a sham operation ( n = 6); however, there was no change in b Ccl4 mRNA expression by qPCR ( n = 3). c Immediately after ON-CR, Nf1 flox/flox ; hGFAP-Cre mice received intraperitoneal injections of anti-CD3 (αCD3) antibodies every other day for 6 weeks. Control mice were injected with anti-IgG antibodies. Optic nerves were analyzed at 12 weeks of age. αCD3 antibody treatment (150 µg) reduced d CD3 + cell content ( n = 5) and e proliferation (%Ki67 + cells; n = 5). f αCD3 antibody treatment following ON-CR does not change Ccl5 RNA expression by qPCR compared to IgG-treated controls (150 µg; n = 3). Increased Ccl5 RNA expression in the optic nerves (qPCR) was observed 7 days after ON-CR in g Nf1 flox/flox ; hGFAP-Cre ( n = 3), h athymic ( Foxn1 nu/nu ; n = 4), and i Rag1 −/− mice ( n = 4) compared to sham controls. Scale bars: a , d , e 50 µm. Two-tailed Student’s t test (ns, not significant)

Article Snippet: Four-week-old Nf1 OPG mice were treated with 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester, Fisher Scientific, 274310), 275 mg/kg PLX3397-containing or control chow pellets (Free Base), 1 mg/ml anti-IL-1β neutralizing antibodies (R&D System 1060-DE-100), anti-IgG2a control antibodies (R&D Systems), or memantine hydrochloride (20 mg/kg) by intraperitoneal injection.

Techniques: Expressing, Control, Injection, RNA Expression, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Brain injury drives optic glioma formation through neuron-glia signaling

doi: 10.1186/s40478-024-01735-w

Figure Lengend Snippet: a Top 3 identified pathways enriched in the optic nerves of Nf1 flox/flox ; hGFAP-Cre mice after optic nerve crush (ON-CR) relative to sham controls following filtering of differentially expressed transcripts from bulk RNA sequencing. b Increased IL-1β RNA expression (qPCR) is observed in the optic nerves of 12-week-old Nf1 flox/flox ; hGFAP-Cre mice following ON-CR at 6 weeks of age relative to sham controls ( n = 4). c Representative IL-1β immunostaining in the optic nerves of Nf1 flox/flox ; hGFAP-Cre mice following ON-CR relative to sham control mice. d RNAscope (in situ RNA hybridization) demonstrates that oligodendrocytes (Olig2 + cells) express IL-1β. e Immunostaining reveals that the majority of the Olig2 + cells co-label with CC1, but not with NG2. f Increasing IL-1β concentrations (0–75 ng/ml) increases microglia Ccl5 protein expression in vitro. g IL-1β-induced microglial Ccl5 production is attenuated by 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester) treatment in vitro ( n = 4). h Anti-IL1β neutralizing antibody treatment (1 mg/ml) of Nf1 flox/flox ; hGFAP-Cre mice immediately after ON-CR at 6 weeks of age results in decreased i optic nerve proliferation (%Ki67 + cells; n = 5) and j Ccl5 mRNA expression (qPCR; n = 3) when analyzed at 12 weeks of age compared to IgG-treated (1 mg/ml) controls. Data are presented as the means ± SEM. Scale bars: c , d , e , i 50 µm. b , c , e , i , j . Two-tailed Student’s t test; f , g One-way ANOVA with Bonferroni post-test correction

Article Snippet: Four-week-old Nf1 OPG mice were treated with 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester, Fisher Scientific, 274310), 275 mg/kg PLX3397-containing or control chow pellets (Free Base), 1 mg/ml anti-IL-1β neutralizing antibodies (R&D System 1060-DE-100), anti-IgG2a control antibodies (R&D Systems), or memantine hydrochloride (20 mg/kg) by intraperitoneal injection.

Techniques: RNA Sequencing, RNA Expression, Immunostaining, Control, RNAscope, In Situ, Hybridization, Expressing, In Vitro, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Brain injury drives optic glioma formation through neuron-glia signaling

doi: 10.1186/s40478-024-01735-w

Figure Lengend Snippet: a Traumatic brain injury (TBI) in Nf1 flox/flox ; hGFAP-Cre mice at 6 weeks of age results in increased b optic nerve volume ( n = 6) and c proliferation (%Ki67 + cells; n = 7) relative to sham treated mice when analyzed at 12 weeks of age, as well as increased TAMs (%Iba1 + cells) and CD3 + T cell content. Increased d Ccl5 RNA expression (qPCR) and e glutamate levels are observed in the optic nerves of Nf1 flox/flox ; hGFAP-Cre mice ( n = 4) 7 days after TBI compared to sham controls. f αIL-1β neutralizing antibody (1 mg/ml) and h memantine treatment (20 mg/kg) both decrease proliferation (%Ki67 + cells; n = 5) and g , i Ccl5 expression ( n = 3) following TBI relative to their respective control mice (IgG and vehicle treatment groups). Data are presented as the means ± SEM. Scale bar: b 100 μm; c , f , i 50 μm. Two-tailed Student’s t test

Article Snippet: Four-week-old Nf1 OPG mice were treated with 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester, Fisher Scientific, 274310), 275 mg/kg PLX3397-containing or control chow pellets (Free Base), 1 mg/ml anti-IL-1β neutralizing antibodies (R&D System 1060-DE-100), anti-IgG2a control antibodies (R&D Systems), or memantine hydrochloride (20 mg/kg) by intraperitoneal injection.

Techniques: RNA Expression, Expressing, Control, Two Tailed Test

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: 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: Cell reports

Article Title: A synaptic molecular dependency network revealed by knockdown of autism- and schizophrenia-associated genes

doi: 10.1016/j.celrep.2023.112430

Figure Lengend Snippet: Key resources table

Article Snippet: anti-Mouse IgG2, goat polyclonal , Novus Biologicals , Cat#NB7513.

Techniques: Recombinant, Gene Expression, Modification, Imaging, Software