Journal: Cell Death & Disease

Article Title: M2 macrophage-mediated interleukin-4 signalling induces myofibroblast phenotype during the progression of benign prostatic hyperplasia

doi: 10.1038/s41419-018-0744-1

Figure Lengend Snippet: a Representative CD68 immunohistochemical staining images in the early-progressed BPH, age-matched prostate and elderly BPH tissues. Scale bar, 200 μm. b Scatter plots showing macrophage clusters in the three analysed groups. The number of macrophage clusters in prostate tissues correlated positively with the stroma-to-epithelium ratio, and percent area densities of α-SMA, collagen I and fibres. c Serial histological sections showing the early-progressed BPH tissues stained for CD68, a macrophage-specific marker, and CD163, an M2 macrophage marker. The distribution of CD68 + and CD163 + cells was almost identical. Scale bar, 200 μm. d Representative immunofluorescence (IF) staining images, showing CD68 (green) and CD163 (red) expression in the infiltrating macrophages in early-progressed BPH tissues. Scale bar, 100 μm. e Quantitative RT-PCR results showing α-SMA, COL1A1 and COL3A1 expression in primary prostate fibroblast (PrPF)-early, PrPF-control and PrPF-old cells co-cultured with THP-1-derived M2 macrophages. Data are shown as relative gene expression compared to that in the respective untreated fibroblasts. f Western blots showing α-SMA and collagen I protein expression in PrPF-early, PrPF-control and PrPF-old cells co-cultured with THP-1-derived M2 macrophages. g Representative IF staining results, showing the co-expression of α-SMA (red) and collagen I (green) following a 48-h co-culture of PrPF-early, PrPF-control and PrPF-old cells with THP-1-derived M2 macrophages. The respective untreated fibroblasts were used as controls. Scale bar, 100 μm. h Solidified collagen-gel shrinkage after the seeding of PrPF-early, PrPF-control and PrPF-old cells, untreated or treated with the conditioned medium. The respective untreated fibroblast samples were used as controls, and all assays were performed in triplicate. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies included mouse monoclonal α-SMA (Abcam, ab7817, 1:100 dilution), CD163 (OriGene, TA506380, 1:200 dilution) and EpCAM (Cell Signaling Technology, #2929, 1:800 dilution) antibodies, and the rabbit primary antibodies included collagen I (Abcam, ab138492, 1:800 dilution), CD68 (Proteintech, 25747-1-AP, 1:400 dilution), IL4R (ImmunoWay, YT2337, 1:400 dilution), TGFβ RI (ImmunoWay, YT4627, 1:400 dilution) and vimentin (Cell Signaling Technology, #5741, 1:100 dilution) antibodies.

Techniques: Immunohistochemical staining, Staining, Marker, Immunofluorescence, Expressing, Quantitative RT-PCR, Control, Cell Culture, Derivative Assay, Gene Expression, Western Blot, Co-Culture Assay

Journal: Human reproduction (Oxford, England)

Article Title: Distribution of decidual natural killer cells and macrophages in the neighbourhood of the trophoblast invasion front: a quantitative evaluation.

doi: 10.1093/humrep/det353

Figure Lengend Snippet: Figure 5 Distribution of dMph. (a) Cryostat section from a cell column at a first trimester placentation site double-stained with an anti-CD163 antibody (red) and an anti-HLA-G antibody (dark blue) as a sample image. Scale bar ¼ 100 mm. (b) Box plots of normalized areal cell densities s of dMph at first trimester placentation sites according to Equation (15), in five trophoblast neighbourhood rings according to Equation (17), are depicted. The dashed line s ¼ 1 represents the average areal cell density. #P ¼ 0.05, ##P ¼ 0.01 compared with 1 by Student’s t-tests (H0:m ¼ 1). ANOVA analysis showed a high significance,P ≤0.01.*P ¼ 0.05,**P ¼ 0.01ofpair-wiseTukeyhonestsignificantdifferenceposthoctests(s ¼ 1).(c)DistributionofdMphinco-culturesof first trimester decidua parietalis with first trimester villous explants using cryostat sections from co-cultures double-stained with an anti-CD163 antibody and an anti-HLA-G antibody. Box plots of normalized areal cell densities s. #P ¼ 0.05, ##P ¼ 0.01 compared with 1 by Student’s t-tests (H0:m ¼ 1). ANOVA analysis showed a high significance, P ≤0.01. **P ¼ 0.01 of pair-wise Tukey honest significant difference post hoc tests (s ¼ 1). Open circle is an outlier. (d) Distribution of dMph in first trimester decidua parietalis determined by using cryostat sections from tissue fragments stained with an anti-CD163 antibody. Box plots of normalized areal cell densities s. P . 0.05 by ANOVA test. Filled circle is an extreme value.

Article Snippet: The following antibodies were used: † monoclonal mouse anti-human CD56 (clone 123C3; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), dilution: 1:400, final concentration: 0.5 mg/ml; † monoclonal mouse anti-human CD163 (clone 5C6-FAT; Acris Antibodies GmbH, Hiddenhausen, Germany), dilution: 1:500, final concentration: 0.4 mg/ml; † monoclonal mouse anti-human HLA-G (clone 4H84; Exbio, Praha, Czech Republic; supplied by Eubio, Vienna, Austria), dilution: 1:500, final concentration: 2 mg/ml.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Fimepinostat Promotes Apoptosis and Decreases Cytokine Secretion in NF2 -Related Human Schwannoma Cells

doi: 10.3390/ijms27062636

Figure Lengend Snippet: Fimepinostat Corrects Loss of Merlin-related Cytokine Expression and Decreases MCP-1 Secretion. ( A , B ). Cytokine array analysis of 10 patient-derived vestibular schwannoma (VS) samples showed IL-6, IL-8, and MCP-1 as the most highly expressed in VS cells alone and VS–monocyte cocultures. ( C ). Addition of monocytes to VS cultures significantly increased secretion of MCP-2, MCP-3, macrophage inflammatory protein-1β (MIP-1b), and osteopontin (OPN) (* p < 0.05). ( D , E ). Immunocytochemistry for CD163+ M2 macrophages (green) and S100B+ VS cells (red), nuclei are stained in blue. ( F ). Cytokine array analysis of HS11 (WT) and HS02 (MD) untreated and treated with 100 nM fimepinostat for 24 h (Mean ± SD, *** p < 0.0005 and **** p < 0.0001). ( G ). MCP-1 ELISA of HS11 and HS02 cells untreated and treated with 100 nM fimepinostat at 6, 16, and 24 h (Mean ± SD, ** p < 0.005 and *** p < 0.0005).

Article Snippet: Immunocytochemistry was performed with anti-CD163 primary antibody (1:100, OriGene, TA506383), S100B (1:100, Abcam, ab52642), AlexaFluor-488- and 594-conjugated secondary antibodies (1:500; Invitrogen), and DAPI nuclear stain, as previously described [ ].

Techniques: Expressing, Derivative Assay, Immunocytochemistry, Staining, Enzyme-linked Immunosorbent Assay