Journal: Advanced Science

Article Title: IL‐4/STAT6‐signaling Influences Local Inflammation and Regeneration Processes During Acute Pancreatitis and Promotes Fibrosis by a Direct Activation of Pancreatic Fibroblasts During Chronic Pancreatitis

doi: 10.1002/advs.202515585

Figure Lengend Snippet: CD206 M2‐like macrophages differentiate in the absence STAT6 but retain proinflammatory properties (A) Immunofluorescent labeling of CD206, CCR2 and CD3 in CP tissue. (B) Quantification of CD206 + , CCR2 + and CD3 + cells in the pancreas of CP mice ( n = 10). (C) Heat map illustrating different levels of inflammation related proteins between wild type and Stat6‐/‐ mice. The numbers in the boxes represents the change compared with untreated control mice, the amount of all red labeled proteins were significantly changed in Stat6‐/‐ compared to wild type mice after onset of CP ( n = 5). (D) Quantitative gene expression analysis by RT‐qPCR of Arg1 , Fizz , Ym1 and Il10 in CP tissue of wild type and Stat6‐/‐ mice ( n = 5). (E) Immunofluorescent labeling of MMP9 (red) and α‐amylase (green) in CP tissue and quantification of MMP9 + cells in the pancreas of CP mice. (F) Quantitative RT‐PCR of Nos2 , Il1b , Mmp9 and Il6 in CP tissue of wild type and Stat6‐/‐ mice ( n = 10). (G) Measurement of TGF‐β in serum of untreated control mice and CP mice ( n = 9). (H) Quantitative RT‐PCR of Tgfbi and Csf1 in CP tissue of wild type and Stat6‐/‐ mice ( n = 5). Data in B and D‐H are shown as mean +/‐ SEM. Differences were tested for statistical significance by unpaired students t‐test or Mann–Whitney for non‐normally distributed samples for independent samples, significance levels of p < 0.05 are marked by an asterisk.

Article Snippet: Recombinant Mouse IL‐4 Protein (#404‐ML, R&D Systems), Recombinant Mouse IL‐13 Protein (#413‐ML, R&D Systems), Lipopolysaccharide Escherichia coli O26:B6 (L8274, Sigma Aldrich), Recombinant Mouse IL‐6 Protein (406‐ML, R&D systems), Recombinant Mouse TGF‐β Protein (7666‐MB, R&D systems), Recombinant mouse IL‐10 (575804, BioLegend).

Techniques: Labeling, Control, Gene Expression, Quantitative RT-PCR, MANN-WHITNEY

Journal: The Journal of Immunology Author Choice

Article Title: NR4A nuclear receptor expression in human macrophages mediates apoptosis and controls Mycobacterium tuberculosis growth

doi: 10.1093/jimmun/vkaf252

Figure Lengend Snippet: Macrophage apoptosis in response to pharmacologic treatment is reduced by NR4A knockdown. (A–C) AML cells were transfected with 15 μM of either scrambled siRNA or siRNA targeting NR4A1, NR4A2, or NR4A3. After 24 h, they were left untreated or treated with 1 μM staurosporine for 8 h, 10 μM actinomycin D for 24 h, or 100 μM etoposide for 24 h. Caspase-3 and caspase-7 cleavage activity was measured by luminescent assay. (D) Representative Western blot showing total and cleaved PARP1 protein expression in AML cells following treatment with staurosporine. (E) Quantified Western blot data are shown from (D). P values are indicated as asterisks above the plots as obtained from one-way ANOVA (* P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001) from n = 3 (A–C) or n = 4 (D, E) unique donors. Data are shown as mean ± SEM.

Article Snippet: Transfected AML cells in 96-well plates were treated with InSolution Staurosporine (Thermo Fisher Scientific), actinomycin D (Tocris Bioscience), or etoposide (Thermo Fisher Scientific) to induce cell death, or infected with M.tb at MOI of 5 for 24 to 72 h. Cell death was assayed in triplicate with the CellTiter Glo Assay (Promega) following the manufacturer’s instructions.

Techniques: Knockdown, Transfection, Activity Assay, Luminescence Assay, Western Blot, Expressing