Journal: Scientific Reports

Article Title: Targeting colony stimulating factor-1 receptor signalling to treat ectopic pregnancy

doi: 10.1038/s41598-020-72785-y

Figure Lengend Snippet: CSF-1R and CSF-1 expression in immortalised first trimester trophoblast cells (SW.71) ( A ) Representative two-dimensional confocal image of SW.71 trophoblast cells immunostained for CSF-1 ligand (green) and nuclei (blue). (B) Three-dimensional cross-section image of the SW.71 cell showing localisation of CSF-1 protein in the cytoplasm (CP). (C) Representative two-dimension confocal image of trophoblast cells immunostained for CSF-1R (green) and nuclei (blue) in the SW.71 cells. (D) Three-dimensional cross-section image of the SW.71 cell showing localisation of CSF-1R in the cell membrane (CM) (Inset) Negative control SW.71 cells with DAPI staining on the nuclei (N) and no immunoreactivity observed. Scale bars A/B/Inset 20 µm and BD 10 µm.

Article Snippet: Blocking was carried out with BSA and normal goat serum for 1 h. CSF-1 and CSF-1R were immunolocalised using primary antibodies conjugated with Alexa Fluor 488 against CSF-1 (Santa Cruz Biotechnology, Texas, USA, sc365779, 1:500) and CSF-1R (Santa Cruz Biotechnology, Texas, USA, sc365719, 1:500) overnight at 4 °C with blocking peptide as a negative control.

Techniques: Expressing, Membrane, Negative Control, Staining

Journal: Scientific Reports

Article Title: Targeting colony stimulating factor-1 receptor signalling to treat ectopic pregnancy

doi: 10.1038/s41598-020-72785-y

Figure Lengend Snippet: Effect of CSF-1 and CSF-1R antagonism with GW2580 on trophoblast cell proliferation, migration and viability (A) Effects of exogenous CSF-1 (50–200 ng/ml) on SW.71 cell proliferation after 48 h exposure. (B) Effects of GW2580 (20 and 40 µM) on proliferation as a percentage of vehicle control after 48 h. (C) Effects of exogenous CSF-1 (100 ng/ml) with GW2580 (5–40 µM) on as a percentage of vehicle control after 48 h. (D) Effect of CSF-1 (100 ng/ml) and GW2580 (20–40 µM) on viable cell number as a percentage of control. (E) The effect of 40 µM GW2580 in the presence or absence of CSF-1 (100 ng/ml) of cell cytotoxicity. (F) A representative image showing scratch imaged at 0 h and at 18 h using TScratch software. (G) Effects of GW2580 (20–40 µM) and exogenous CSF-1 ligand (100 ng/ml) on SW.71 cellular migration after an 18 h incubation. Vehicle (DMSO) treated cells were used as controls and values normalised to the control. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Blocking was carried out with BSA and normal goat serum for 1 h. CSF-1 and CSF-1R were immunolocalised using primary antibodies conjugated with Alexa Fluor 488 against CSF-1 (Santa Cruz Biotechnology, Texas, USA, sc365779, 1:500) and CSF-1R (Santa Cruz Biotechnology, Texas, USA, sc365719, 1:500) overnight at 4 °C with blocking peptide as a negative control.

Techniques: Migration, Control, Software, Incubation

Journal: Scientific Reports

Article Title: Targeting colony stimulating factor-1 receptor signalling to treat ectopic pregnancy

doi: 10.1038/s41598-020-72785-y

Figure Lengend Snippet: Immunolocalisation of CSF-1R and CSF-1 in tubal EP implantation sites. (A ) Representative haematoxylin and eosin image of the ectopic implantation site in the Fallopian tube. (B) Negative control DAPI stained section (blue) showing no CSF-1R staining (red). (C) Section of human Fallopian tube ectopic implantation site immunostained for CSF-1R showing staining (red) and confirming the expression of CSF-1R. (D , E) Representative image showing immunolocalisation of CSF-1 in trophoblast invading a human FT. Immunolocalisation of CSF-1 at ectopic implantation sites is observed as green staining. (F) CSF-1 immunostaining (green staining highlighted by white arrows) with DAPI (blue) staining the nuclei of trophoblasts at ectopic implantation site. (G) negative control section with DAPI (blue) staining. (H , I) Representative image showing immunolocalisation (green) of CSF-1R in trophoblast invading a human FT. (J) Immunolocalisation of CSF-1R on ectopic implantation sites is shown by green staining (white arrows) with DAPI (blue) staining nuclei of trophoblasts at ectopic implantation site. (K) No immunostaining in control section with DAPIU (blue staining). FT Fallopian tube, CTB cytotrophoblast, STB syncytiotrophoblast. Scale bars represent 100 µm.

Article Snippet: Blocking was carried out with BSA and normal goat serum for 1 h. CSF-1 and CSF-1R were immunolocalised using primary antibodies conjugated with Alexa Fluor 488 against CSF-1 (Santa Cruz Biotechnology, Texas, USA, sc365779, 1:500) and CSF-1R (Santa Cruz Biotechnology, Texas, USA, sc365719, 1:500) overnight at 4 °C with blocking peptide as a negative control.

Techniques: Negative Control, Staining, Expressing, Immunostaining, Control

Journal: PLoS ONE

Article Title: Vascularization and biocompatibility of poly(ε-caprolactone) fiber mats for rotator cuff tear repair

doi: 10.1371/journal.pone.0227563

Figure Lengend Snippet: Results from quantification of immunofluorescent detection of immune cell markers.

Article Snippet: For detection of capillaries, endothelial cells were immunohistochemically stained using a rabbit anti-mouse CD31 antibody (LifeSpan Biosciences, Seattle, USA purchased from BIOZOL Diagnostica Vertrieb GmbH, Eching, Germany); a rat anti-mouse CD68 antibody (Acris Antibodies GmbH, Herford, Germany) was used to detect macrophages; a rabbit anti-mouse CSF1R antibody (OriGene Technologies, Inc., Rockville, USA) was used for detection of macrophages in general; a rat anti-mouse CD86 antibody (OriGene Technologies, Inc., Rockville, USA) was used to detect macrophages with predominantly pro-inflammatory actions; a rabbit anti-mouse CD11b antibody (OriGene Technologies, Inc., Rockville, USA) was used to detect monocytes and a rat anti-mouse CD3 antibody (OriGene Technologies, Rockville, Inc., USA) was used to detect T-cells.

Techniques:

Journal: PLoS ONE

Article Title: Vascularization and biocompatibility of poly(ε-caprolactone) fiber mats for rotator cuff tear repair

doi: 10.1371/journal.pone.0227563

Figure Lengend Snippet: Representative histological stainings for detection of macrophages in general, macrophages with predominantly pro-inflammatory actions, monocytes and T-cells (all green fluorescence) (A-C, E-G, I-K, M-O) 14 days after implantation into the dorsal skinfold chamber of BALB/c mice. (A-C) Immunofluorescence detection of CSF1R for detection of macrophages in general. (E-G) Immunofluorescence detection of CD86 for detection of macrophages with predominantly pro-inflammatory actions. (I-K) Immunofluorescence detection of CD11b for detection of monocytes. (M-O) Immunofluorescence detection of CD3 for detection of T-cells. Nuclei were stained using DAPI (blue fluorescence). Scale bars: 50 μm. (D,H,L,P) Quantitative analysis of cell infiltration by immunofluorescent staining for CSF1R (D) , CD86 (H) , CD11b (L) and CD3 (P) . All data are expressed as the fluorescence intensity of the marker of interest normalized to the fluorescence intensity of DAPI staining. Values represent means ± SEM; *p < 0.05 vs. control and PCL; + p < 0.05 vs. control and CS-g-PCL.

Article Snippet: For detection of capillaries, endothelial cells were immunohistochemically stained using a rabbit anti-mouse CD31 antibody (LifeSpan Biosciences, Seattle, USA purchased from BIOZOL Diagnostica Vertrieb GmbH, Eching, Germany); a rat anti-mouse CD68 antibody (Acris Antibodies GmbH, Herford, Germany) was used to detect macrophages; a rabbit anti-mouse CSF1R antibody (OriGene Technologies, Inc., Rockville, USA) was used for detection of macrophages in general; a rat anti-mouse CD86 antibody (OriGene Technologies, Inc., Rockville, USA) was used to detect macrophages with predominantly pro-inflammatory actions; a rabbit anti-mouse CD11b antibody (OriGene Technologies, Inc., Rockville, USA) was used to detect monocytes and a rat anti-mouse CD3 antibody (OriGene Technologies, Rockville, Inc., USA) was used to detect T-cells.

Techniques: Fluorescence, Immunofluorescence, Staining, Marker, Control

Journal: bioRxiv

Article Title: Crosstalk between Stromal cells and Macrophages Shapes Host Immunity to Mycobacteria

doi: 10.64898/2026.02.05.704052

Figure Lengend Snippet: A. Gating strategy depicting differences in macrophage subsets in the peritoneal lavage in control and BCG infected mice. Initial plot was pre-gated on CD115 + CD45 + single cells. B. Depicted is mean ± SEM of SPM and LPM in control mice (n=11), BCG-infected mice 2 weeks p.i. (n=6), 4 weeks p.i. (n=6), 6 weeks p.i. (n=5) in 10 independent experiments. 2 out of 12 experiments were removed, where insufficient i.p. infection was suspected. Two-way ANOVA with Dunnett’s multiple comparisons test. C. Bar shows mean ± SEM of BFP-positive LPM 30 min post injection of BCG-BFP (n=3 mice of 3 independent experiments). D. Depicted are means ± SEM of neutrophils, Ly6C + monocytes and LPM in PL 24h p.i. of BCG (n=4) compared to control mice (n=4) in 2 independent experiments (two-way ANOVA). E. Graphical abstract of adoptive LPM transfer. F. Immunofluorescence staining for podoplanin AF647 and F4/80 PE of an omentum whole mount of an infected mouse post LPM transfer (compare ). The staining was performed in 5 independent mice. G. Depicted is the mean ± SEM of percentage of LPM and transferred GFP + LPM in peritoneal lavage 24h p.t. in PBS (n=6) and BCG-infected mice (n=7) in 7 independent experiments. One PBS mouse was removed due to an unrelated abdominal pathology (hydronephrosis). H. Shown is mean ± SEM of the relation of GFP-positive transferred cells in omentum (om) and PL (lav) relative to single cells (singlets), (unpaired t-test, compare ). I. Depicted are means ± SEM of neutrophils and Ly6C + monocytes in PL 2 (n=4) and 4 weeks (n=6) p.i. compared to control (n=7) in 7 independent experiments (Brown-Forsythe and Welch ANOVA). 1 mouse was removed, where insufficient i.p. infection was suspected. J. iNOS staining was performed in 6 experiments of 3I. Depicted is mean ± SEM (Two-way ANOVA). K. Depicted are means ± SEM of immune cell populations in PL of control (n=11) and infected (n=12) WT and CXCR4 creER :R26 tdTomato mice 16 weeks post infection (8 independent experiments). L. Exemplary FACS plots of macrophage subsets in PL in control mice and 16 weeks p.i. with BCG. M. Depicted is mean ± SEM of tomato-positive cells in the MHC-II + F4/80 + population from Cxcr4 creER :R26 tdTomato mice 16 weeks p.i., if detected (n=5 in 3 experiments). In 1 infected mouse no double positive cells were found. N. Peritoneal macrophage populations with and without antibiotic treatment (n=4 in 3 independent experiments, Two-way ANOVA with Sidak multiple comparisons test). O. Depicted are means ± SEM of small and large pleural macrophages and the percentage of tomato-positive cells in Cxcr4 creER :R26 tdtomato mice 6 weeks post induction for controls (n=6) and 2w-i.p.-infected mice (n=6, in 6 experiments) (left) and mice 20 weeks post induction for controls (n=5) and 16w-i.p.-infected mice (n=6, in 3 experiments (right). The mouse excluded in 1J was also excluded here. P. Graphs show means ± SEM of pleural macrophage populations in control (n=5) and M.tb lung infected mice (n=9) in two independent experiments (Welch’s unpaired t-test).

Article Snippet: CD115 PE (AFS98, #130-122-018), CD31 FITC (390, #1301-236-75), Ter119 Biotin (Ter119, #130-120-828), CD3e Biotin (145-2C11, #130-101-990), CD19 Biotin (6D5, #130-119-797), SiglecF Biotin (ES22-10D8, #130-119-136), Sca-1 Biotin (D7, #130-126-949), CD127 Biotin (A7R34 and REA680, #130-102-039 and #130-110-373), CD115 PE REAlease (REAL272, #130-118-071) and CD115 Antibody Kit, PE, REAlease® (#130-122-632) were bought from Miltenyi Biotec and CD11c FITC (HL3, #553801), TNF APC (MP6-XT22; #554420), Ly6G PE (1A8, #551461), CD103 PE (M290, #561043) from BD bioscience.

Techniques: Control, Infection, Injection, Immunofluorescence, Staining