Journal: Journal of Inflammation Research

Article Title: Regulation of the Immune Microenvironment by an NLRP3 Inhibitor Contributes to Attenuation of Acute Right Ventricular Failure in Rats with Pulmonary Arterial Hypertension

doi: 10.2147/JIR.S336964

Figure Lengend Snippet: Flow Cytometry Antibody

Article Snippet: CD45 , 561586 , Rat CD45 APC-Cy7 OX-1 50ug , BD Pharmingen.

Techniques: Flow Cytometry, Marker

Journal: Journal of Inflammation Research

Article Title: Regulation of the Immune Microenvironment by an NLRP3 Inhibitor Contributes to Attenuation of Acute Right Ventricular Failure in Rats with Pulmonary Arterial Hypertension

doi: 10.2147/JIR.S336964

Figure Lengend Snippet: Changes in the proportion of immune cells in myocardial tissue were analysed via flow cytometry. ( A ) The proportion of CD45 + cells in each group was detected by flow cytometry. ( B ) The proportion of CD45 + CD11b + cells in each group was detected by flow cytometry. ( C ) The proportion of CD68 + CD86 + cells in each group was detected by flow cytometry. ( D ) Pulmonary hypertension and LPS caused a significant increase in the proportion of CD45-positive inflammatory cells in right ventricular tissue (normal vs PAH, normal vs normal + LPS, n = 3). ( E ) CD45-positive/CD11b-positive mononuclear macrophage infiltration was significantly elevated in right ventricular tissues from rats subjected to PAH or LPS (normal vs PAH, normal vs normal + LPS, normal + LPS vs PAH + LPS, n = 3). ( F ) The proportion of CD68-positive/CD86-positive M1 macrophages was decreased in the hearts of PAH rats and significantly increased in the right ventricular tissues of PAH rats stimulated with LPS, but MCC950 application significantly inhibited the change in the proportion of CD68-positive/CD86-positive M1 macrophages (n = 3). The significance of the difference was analysed by one-way ANOVA, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: CD45 , 561586 , Rat CD45 APC-Cy7 OX-1 50ug , BD Pharmingen.

Techniques: Flow Cytometry

Journal: Cell reports

Article Title: RAG-Mediated DNA Breaks Attenuate PU.1 Activity in Early B Cells through Activation of a SPIC-BCLAF1 Complex

doi: 10.1016/j.celrep.2019.09.026

Figure Lengend Snippet: (A-C) Art −/− :μIgh:Bcl2 pre-B cells were transduced with a retrovirus expressing a scrambled shRNA (−) or shBclaf1 (+) and then subsequently withdrawn from IL-7. (A and B) Spic and Syk mRNA expression assessed in indicated small pre-B cells 2 days after IL-7 withdrawal. Data are mean and SE for three independent experiments. (C) Western blot of SYK and BCLAF1 in indicated small pre-B cells 2 days after IL-7 withdrawal. Data are representative of three independent experiments. (D) Flow cytometric analysis showing EGFP (y axis) and FSC (x axis) in bone marrow pre-B cells (B220 lo CD43 − IgM − ) from wild-type and Spic igfp / igfp mice. Data are representative of five independent experiments. (E) Percentage of EGFP-positive small pre-B cells in Spic igfp / igfp (circles) and Atm −/− :Spic igfp / igfp (squares) mice was quantified by flow cytometry as in (D). Data are mean and SE from three independent mice of each genotype. (F–H) Syk mRNA expression (F), ChIP-PCR of PU.1 at Syk promoter (G), and ChIP-PCR of BCLAF1 at Syk promoter (H) in EGFP-negative (−) and EGFP-expressing (+) small pre-B cells sorted from Spic igfp / igfp mice. Data in (F) are the mean and SE from three independent experiments. Data in (G) and (H) are representative of two independent experiments. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001,****p ≤ 0.0001; ns, not significant.

Article Snippet: PE-Cy7-conjugated anti-CD45/B220 (clone RA3–6B2) , BD Biosciences , Cat # 552772: RRID: AB_394458.

Techniques: Transduction, Expressing, shRNA, Western Blot, Flow Cytometry

Journal: Cell reports

Article Title: RAG-Mediated DNA Breaks Attenuate PU.1 Activity in Early B Cells through Activation of a SPIC-BCLAF1 Complex

doi: 10.1016/j.celrep.2019.09.026

Figure Lengend Snippet: (A) Western blot of BCLAF1 in sorted CD19− (non-B cell) and CD19+ B cell populations from bone marrow of 5-week-old Bclaf1 f/f :Mb1-cre mice. Data are representative of three independent mice. (B) Flow cytometric analysis of BrdU incorporation (y axis) and DNA content (7AAD, x axis) performed 24 h after IL-7 withdrawal. Percentage of cells that entered S phase during BrdU labeling (box) is indicated. Data are representative of at least three independent experiments. (C) Percentage of cells that entered S phase in cell cycle analysis performed in (B). Data are mean and SE for four independent experiments. (D) Syk mRNA expression 24 h after IL-7 withdrawal. Data are mean and SE for three independent experiments. (E) Quantitation of flow cytometric analysis of pro-B cells (B220 lo IgM − CD43 + ) and pre-B cells (B220 lo IgM − CD43 − ) in bone marrow of 5-week-old Bclaf1 f/f (black bars, n = 12), Mb1-cre (gray bars, n = 9), and Bclaf1 f/f :Mb1-cre (white bars, n = 12) mice. Large and small pre-B cells were gated on the basis of forward-scatter and side-scatter characteristics. (F) Syk mRNA expression in small and large pre-B cells sorted from 5-week-old Bclaf1 f/f (black bars, n = 4), Mb1-cre (gray bars, n = 5), and Bclaf1 f/f :Mb1-cre (white bars, n = 5) mice. Data in (E) and (F) are mean and SE for indicated numbers of mice. *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001; ns, not significant. See also Figure S6 .

Article Snippet: PE-Cy7-conjugated anti-CD45/B220 (clone RA3–6B2) , BD Biosciences , Cat # 552772: RRID: AB_394458.

Techniques: Western Blot, BrdU Incorporation Assay, Labeling, Cell Cycle Assay, Expressing, Quantitation Assay

Journal: Cell reports

Article Title: RAG-Mediated DNA Breaks Attenuate PU.1 Activity in Early B Cells through Activation of a SPIC-BCLAF1 Complex

doi: 10.1016/j.celrep.2019.09.026

Figure Lengend Snippet:

Article Snippet: PE-Cy7-conjugated anti-CD45/B220 (clone RA3–6B2) , BD Biosciences , Cat # 552772: RRID: AB_394458.

Techniques: Recombinant, SYBR Green Assay, Purification, Magnetic Beads, Plasmid Preparation, Software

Journal: Journal of Crohn's & Colitis

Article Title: Cannabinoid Receptor Activation on Haematopoietic Cells and Enterocytes Protects against Colitis

doi: 10.1093/ecco-jcc/jjaa253

Figure Lengend Snippet: List of reagents.

Article Snippet: APC/Cy7 anti-mouse CD45 , 30-F11 , 103115 , Biolegend.

Techniques: Recombinant, Purification

Journal: Journal of Crohn's & Colitis

Article Title: Cannabinoid Receptor Activation on Haematopoietic Cells and Enterocytes Protects against Colitis

doi: 10.1093/ecco-jcc/jjaa253

Figure Lengend Snippet: Cannabinoids ameliorate TNBS-induced colitis and reduce effector cell phenotypes. Female BALB/c mice were injected intrarectally with 100 mg/kg TNBS in 50% ethanol. Starting 3 days before disease induction and continuing daily, mice were gavaged with either: vehicle [10% EtOH in PBS+Tween-80], CBD [10 mg/kg], THC [10 mg/kg], or a combination of THC and CBD [10 mg/kg, both], [n = 10]. Mice were sacrificed at 5 days post disease induction, and blood as well as organs of interest were harvested and analysed for colitis-relevant parameters. [A] Percent weight change and [B] actual weight change over the course of disease. [C] Representative colonoscopy images taken on Day 5. [D] Quantification of colitis scores at indicated time points throughout disease course, [n = 5 per group, per time point]. [E] Representative image and [F] length of colons at sacrifice [n = 10]. [G‐I] ELISAs from serum at sacrifice quantifying disease-relevant biomarkers of colitis severity [n = 4–5]. [J] PAS stain of proximal colons from representative mice taken at sacrifice. [K] Representative flow cytometry psuedocolour dot plots [gate: Live,CD45+] displaying effector cell types in the cLP [n = 6]. [L] Offset histograms of FoxP3 expression [gate: Live, CD45+CD4+] in cLP [n = 6]. Each symbol represents an individual mouse. Data are presented as mean ± standard error of the mean [SEM]. NS, not significant; *p <0.05; **p <0.01, ***p <0.001, ****p <0.0001 by two-way ANOVA with Tukey’s multiple comparisons test. Data are from one experiment representative of three independent experiments.

Article Snippet: APC/Cy7 anti-mouse CD45 , 30-F11 , 103115 , Biolegend.

Techniques: Injection, Staining, Flow Cytometry, Expressing

Journal: Journal of Crohn's & Colitis

Article Title: Cannabinoid Receptor Activation on Haematopoietic Cells and Enterocytes Protects against Colitis

doi: 10.1093/ecco-jcc/jjaa253

Figure Lengend Snippet: Cannabinoids prevent DSS-induced colitis and reduce effector cell phenotypes. Female C57BL/6 mice were treated with either: vehicle [10% EtOH in PBS+Tween-80], CBD [10 mg/kg], THC [10 mg/kg], or a combination of THC and CBD [10 mg/kg, both] by oral gavage for 3 days before 2% DSS was added to their drinking water. DSS remained in the drinking water until termination of the study 14 days later. Treatments continued daily. Mice were sacrificed at 14 days post disease induction, and blood as well as organs of interest were harvested and analysed for colitis-relevant parameters. [A] Percent weight change and [B] stool score assessed over the course of disease [n = 5]. [C] Representative colonoscopy images taken on Day 10. [D] Quantification of colitis scores at indicated time points throughout disease course [n = 8 per group, per time point]. [E] Representative image and [F] length of colons at sacrifice [n = 5]. [G‐I] ELISAs from serum at sacrifice quantifying disease relevant biomarkers of colitis severity [n = 10, SAA n = 5, LCN-2, MPO]. [J] PAS stain of proximal colons from representative mice taken at sacrifice. [K] Representative flow cytometry psuedocolour dot plots [gate: Live,CD45+] displaying effector cell types in the cLP. [L] Representative flow cytometry contour plots [gate: Live,CD45+CD3+CD4+] of T-bet+ Th1 or Gata3+ Th2 cells in the cLP. Each symbol represents an individual mouse. Data are presented as mean ± standard error of the mean [SEM]. NS, not significant; *p <0.05; **p <0.01, ***p <0.001, ****p <0.0001 by two-way ANOVA with Tukey’s multiple comparisons test. Data are from one experiment representative of four independent experiments.

Article Snippet: APC/Cy7 anti-mouse CD45 , 30-F11 , 103115 , Biolegend.

Techniques: Staining, Flow Cytometry