Journal: Science Advances

Article Title: The tumor suppressor adenomatous polyposis coli regulates T lymphocyte migration

doi: 10.1126/sciadv.abl5942

Figure Lengend Snippet: Primary and secondary antibodies/reagents used for immunofluorescence.

Article Snippet: APC anti-human CD11a , Mouse IgG1 , Miltenyi Biotec 130-127-294 , 1:50.

Techniques: Immunofluorescence, Purification

Journal: Science Advances

Article Title: The tumor suppressor adenomatous polyposis coli regulates T lymphocyte migration

doi: 10.1126/sciadv.abl5942

Figure Lengend Snippet: Antibodies used for flow cytometry.

Article Snippet: APC anti-human CD11a , Mouse IgG1 , Miltenyi Biotec 130-127-294 , 1:50.

Techniques: Cytometry

Journal: Science Advances

Article Title: The tumor suppressor adenomatous polyposis coli regulates T lymphocyte migration

doi: 10.1126/sciadv.abl5942

Figure Lengend Snippet: Antibodies used for Western blot.

Article Snippet: APC anti-human CD11a , Mouse IgG1 , Miltenyi Biotec 130-127-294 , 1:50.

Techniques: Western Blot

Journal: Journal of Crohn's & Colitis

Article Title: Intestinal Epithelial Cells Express Immunomodulatory ISG15 During Active Ulcerative Colitis and Crohn’s Disease

doi: 10.1093/ecco-jcc/jjaa022

Figure Lengend Snippet: Expression of ISGs in colonic epithelium during active IBD. [A] RNASeq data of ISGs in laser-captured, microdissected epithelial monolayer [ n = 29]. Heat map shows log2 ratios of gene expression in microdissected epithelial cells from inflamed, active IBD [ n = 12] compared with expression in epithelial cells from uninflamed IBD and normal controls [ n = 17]. Differential gene expression analysis was performed as described in the Methods section 2.9; *adjusted p -values <0.05. The gene lists are shown in , available as Supplementary data at ECCO-JCC online. [B–E] Expression of pSTAT, CD11a, and ISG15 [brown] in colonic mucosa from active UC and CD compared with non-IBD control [representative pictures from n = 3–4 in each group]. Insets show higher magnification of areas as indicated. Scale bars 100 µm in [B–D], 50 µm in [E]. IHC showing that in active IBD there are [B] increased expression of pSTAT1 in both epithelial cells and immune cells infiltrating the lamina propria, [C] infiltration of CD11a [gene name ITGAL ] expressing immune cells in in the full depth of lamina propria, and [D] increased expression of ISG15 in particularly the surface epithelium. [E] ISH of ISG15 mRNA confirming increased expression in colonic epithelial cells during active IBD. [F–H] Scatter plot and Spearman rank correlation analysis of ISG15 mRNA expression versus IFN γ [F], ITGAL [G], and ITGB2 [H] mRNA expression in colonic pinch biopsies from individual non-IBD controls [NC] [ n = 20], inactive IBDi [ n = 65] and active IBDa [ n = 49] [total n = 134]. IBD, inflammatory bowel disease; UC, ulcerative colitis; CD, Crohn’s disease; IHC, immunohistochemistry; ISH, in situ hybridisation.

Article Snippet: For real-time quantitative polymerase chain reaction [RT-qPCR] of ITGAL and ITGB2 expression in PBMCs [Sample set II], RNA extraction and reverse transcription were done as previously desrcibed. cDNA was analysed for LFA-1 subunits ITGAL and ITGB2 transcripts by quantitative real-time PCR using Taqman probes [ ITGAL : probe ID Hs_00158218_m1; ITGB2 : probe ID Hs_00164957_m1] with the reference genes beta actin [ACTB: probe ID Hs01060665_g1], TATA box binding protein [TBP: probe ID Hs00427620_m1] and eukaryotic 18S rRNA [18S: probe ID Hs99999901_s1] [Applied Biosystems].

Techniques: Expressing, Gene Expression, Control, Immunohistochemistry, In Situ, Hybridization

Journal: Journal of Crohn's & Colitis

Article Title: Intestinal Epithelial Cells Express Immunomodulatory ISG15 During Active Ulcerative Colitis and Crohn’s Disease

doi: 10.1093/ecco-jcc/jjaa022

Figure Lengend Snippet: ISG15 and IL12 regulate IFNγ release from PBMCs. [A] IFNγ release from PBMCs was detected by ELISA after treatment with ISG15 [500 ng/ml], IL12 [20 ng/ml], or ISG15 + IL12 [500 + 20 ng/ml] for 48 h. The plot shows mean + SD IFNγ release from three healthy donors with independent experiments marked as dots [ n = 4]. [B] ISG15-regulated enhanced release of IFNγ from PBMCs isolated from non-IBD controls [ n = 6], inactive IBD [ n = 16], and active IBD [ n = 13]. The plot shows ratio IFNγ release [mean + SEM] between ISG15 + IL12 and IL12 alone in the three groups, with individuals indicated by dots. [C] No significant correlation between PBMC subpopulations and ratio IFNγ release from PBMCs after treatment with IL12 + ISG15 versus IL12 alone. The plot show PBMC subpopulations in 35 samples [Sample set II]. FOP: frequency of parental, CD4+ [T helper cells, monocytes, macrophages, dendritic cells], CD8+ [cytotoxic T cells], CD16+ [natural killer cells, monocytes, and macrophages], CD19+ [B cells], CD14+ [co-receptor, PRR]. [D] Expression of ISG15 receptor units, ITGAL and ITGB2 mRNAs in PBMCs from non-IBD controls [ n = 7], inactive IBD [ n = 17], and active IBD [ n = 15] quantified by RT-qPCR. Fold changes were calculated using the ΔΔCT method, where individual expression levels were determined relative to the mean of reference genes and further relative to the mean in the healthy non-IBD control group. The figures show mean expression +SD and individual fold changes determined relative to the reference gene [ beta actin ] for each group. [E] Correlation between fold change levels of ISG15 receptor units [ ITGB2 and ITGAL ] mRNAs and ratio IFNγ release from PBMCs after treatment with IL12 + ISG15 versus IL12 alone. Pearson correlation coefficients and p -values are shown; p -values for each group in A and D were calculated with one-way ANOVA followed by Dunnett’s multiple comparison test; * p <0.05, **** p <0.0001. IBD, inflammatory bowel disease; PMBC, peripheral blood mononuclear cells; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation; SEM, standard error of the mean; ANOVA, analysis of variance; PRR, pattern recognition receptor; RT-qPCR, real-time quantitative polymerase chain reaction.

Article Snippet: For real-time quantitative polymerase chain reaction [RT-qPCR] of ITGAL and ITGB2 expression in PBMCs [Sample set II], RNA extraction and reverse transcription were done as previously desrcibed. cDNA was analysed for LFA-1 subunits ITGAL and ITGB2 transcripts by quantitative real-time PCR using Taqman probes [ ITGAL : probe ID Hs_00158218_m1; ITGB2 : probe ID Hs_00164957_m1] with the reference genes beta actin [ACTB: probe ID Hs01060665_g1], TATA box binding protein [TBP: probe ID Hs00427620_m1] and eukaryotic 18S rRNA [18S: probe ID Hs99999901_s1] [Applied Biosystems].

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Expressing, Quantitative RT-PCR, Control, Comparison, Standard Deviation, Real-time Polymerase Chain Reaction

Journal: Particle and Fibre Toxicology

Article Title: Controlled human wood smoke exposure: oxidative stress, inflammation and microvascular function

doi: 10.1186/1743-8977-9-7

Figure Lengend Snippet: Mean ± SEM of gene expression (mRNA levels per 10 6 18 S mRNA) and the geometric mean ± SEM of adhesion molecule expression by FACS analysis in peripheral blood mononuclear cells (PBMCs) at four times after each of the three exposure scenarios

Article Snippet: The assay IDs for the genes were as follows: ITGAL , Hs01035619_m1; CCL2 , Hs00234140_m1; IL6 , Hs00985641_m1; IL8 , Hs00174103_m1; TNF , Hs00174128_m1; HMOX1 , Hs00157965_m1; OGG1 , Hs01114116_gl.

Techniques: Gene Expression, Expressing