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: Cell Reports

Article Title: De novo steroidogenesis in tumor cells drives bone metastasis and osteoclastogenesis

doi: 10.1016/j.celrep.2024.113936

Figure Lengend Snippet: Pregnenolone promotes human osteoclastogenesis and is secreted by human tumor cells (A–C) Representative images (A) and quantification (B and C) of TRAP-stained cell cultures of human blood mononuclear cell-derived OCs cultured for 2 days in the presence of 20 ng/mL recombinant human M-CSF and 20 ng/mL soluble human RANKL and then in the presence of the indicated concentration of pregnenolone or vehicle for 12 days. Scale bars represent 100 μm. (D) Levels of pregnenolone in the supernatants of A375, MDA-MB-435S, A549, NCI-H1299, NCI-H460, MCF-7, MDA-MB-231, DU145, LNCaP, PC-3, Hs895.T, SK-MEL-28, BEAS-2B, NCI-H128, NCI-H2126, HCC70, MCF-10A, MDA-PCa-2b, RWPE-1, and LASCPC-01 cells measured by ELISA. (E) Kaplan-Meier curve of breast-cancer-specific survival for 2,976 patients with high or low expression of Cyp11a1 in the primary tumor. Bar graphs show mean and SD of data from 3 independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.002; n.s., not significant.

Article Snippet: Mouse Competitive ELISA Kit M-CSF , Elabscience Biotech. , Cat# E-EL-M2445.

Techniques: Staining, Derivative Assay, Cell Culture, Recombinant, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Cell Reports

Article Title: De novo steroidogenesis in tumor cells drives bone metastasis and osteoclastogenesis

doi: 10.1016/j.celrep.2024.113936

Figure Lengend Snippet:

Article Snippet: Mouse Competitive ELISA Kit M-CSF , Elabscience Biotech. , Cat# E-EL-M2445.

Techniques: Virus, shRNA, Recombinant, Modification, Staining, RNA Library Preparation, TaqMan Assay, Competitive ELISA, RNA Sequencing, Plasmid Preparation, Software, Real-time Polymerase Chain Reaction

Journal: Oncotarget

Article Title: miR-1207-5p suppresses lung cancer growth and metastasis by targeting CSF1.

doi: 10.18632/oncotarget.8718

Figure Lengend Snippet: Figure 1: CSF1 is a direct target of miR-1207-5p. (A) Left: Scheme for the potential binding sites of miR-1207-5p in the 3ʹ-UTR of CSF1, and the sequence of each intact miR-1207-5p binding site (wild-type, WT) and its mutant (Mut) within the luciferase reporter vector. Right: HEK-293 cells were cotransfected with the miR-1207-5p mimics (or control) and a luciferase reporter containing the 3ʹ-UTR (WT or Mut) of CSF1 for 36 hrs. Cell lysates were then harvested for luciferase activities assay. (B) HEK293, A549, H358 and HK-1 cells were transfected with either miR-1207-5p mimics or mimics control for 48 hrs, the CSF1 mRNA levels were then analyzed by RT–qPCR. (C) RT-qPCR analysis of the expression levels of CSF1 and miR-1207-5p in multiple cancer cell lines. (D–E) A549 cells were transfected with either miR-1207-5p mimics or inhibitors for 48 hrs. CSF1 protein levels in cell lysates were analyzed by Western blotting (D), and in cell supernatant were analyzed by ELISA (E). (F) A549 cells were transfected with indicated vectors and mimics for 48 hrs. Flag-CSF1 mRNA and protein levels were analyzed by RT–qPCR (left) or Western blotting (right). All data are shown as means ± s.e.m. *P < 0.05, **P < 0.01 compared with control.

Article Snippet: The CDS and CDS plus 3ʹUTR of CSF1 were obtained by PCR from CSF1 plasmid (Origene, Rockville, MD, USA) and subcloned into the pCMV-NFlag vector (Beyotime Biotechnology, Jiangsu, China).

Techniques: Binding Assay, Sequencing, Mutagenesis, Luciferase, Plasmid Preparation, Control, Transfection, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: miR-1207-5p suppresses lung cancer growth and metastasis by targeting CSF1.

doi: 10.18632/oncotarget.8718

Figure Lengend Snippet: Figure 4: miR-1207-5p suppresses the metastasis of A549 cells in a nude mouse xenograft model. The tail veins of nude mice were intravenously injected with A549-luciferase cells (which had been treated with either miR-1207-5p agomir or a control agomir for 24 hrs). miR-1207-5p agomir group has nine mice and control agomir group has ten mice. After 7 days, each group of nude mice were treated with miR-1207-5p agomir or control agomir through vein injection, respectively. Fifty days later, D-Luciferin was administered to each mouse, and the mice were then imaged by using a Xenogen IVIS Lumina II imaging system. After then, necropsies and pathology examination were performed. (A) Representative images of IVIS imaging, which revealed the metastatic nodules in the lung (left). Effects of miR- 1207-5p agomir on miR-1207-5p and CSF1 mRNA expression in A549 cells assayed by RT-qPCR (right). (B) Representative images of H&E– stained mice lung sections, which revealed the metastatic nodules (left), and numbers of metastases in lungs of individual mice were analyzed (right). (C) The expression of CSF1, p-AKT and p-STAT3 were assayed by immunohistochemistry (left). The mRNA levels of miR-1207-5p and CSF1 of mice lung tumor tissues were analyzed by RT-qPCR (right). Scale bar: 50 μm. All data are shown as the mean ± s.e.m. *p < 0.05, **p < 0.01 compared with control.

Article Snippet: The CDS and CDS plus 3ʹUTR of CSF1 were obtained by PCR from CSF1 plasmid (Origene, Rockville, MD, USA) and subcloned into the pCMV-NFlag vector (Beyotime Biotechnology, Jiangsu, China).

Techniques: Injection, Luciferase, Control, Imaging, Expressing, Quantitative RT-PCR, Staining, Immunohistochemistry

Journal: Oncotarget

Article Title: miR-1207-5p suppresses lung cancer growth and metastasis by targeting CSF1.

doi: 10.18632/oncotarget.8718

Figure Lengend Snippet: Figure 5: Expression of miR-1207-5p and CSF1 in NSCLC and control non-cancerous lung tissues by means of TMA. (A) In situ hybridization staining of miR-1207-5p mRNA in adenocarcinoma and squamous carcinoma tissues as well as non-cancerous lung tissues (upper panel). Immunohistochemical staining of CSF1 proteins in lung cancer tissues (with or with out lymph node metastasis) and control non-cancerous lung tissues (bottom panel). Scale bar: 50 μm. (B) Expression scores of miR-1207-5p and CSF1 in lung cancer tissues (n = 289) and non-cancerous lung tissues (n = 53). (C) Expression scores of miR-1207-5p and CSF1 in lung cancer tissues, which had both primary tumor tissues and metastasis tumor tissues (n = 118 pairs). All data are shown as the mean ± s.e.m. *p < 0.05, **p < 0.01, ***P < 0.001 compared with control. (D) Kaplan-Meier analysis was used to plot the overall survival curves of 289 cases of NSCLC patients with different expression of miR-1207-5p and CSF1, which statistical significance was assessed by log-rank test (left and middle panel). MIRUMIR database [24] was used to analyze the clinic impact of CSF1 expression patterns on lung cancer patients’ survival in a NSCLC cancer specimen expression profile dataset (GSE31210, n = 222) (right panel).

Article Snippet: The CDS and CDS plus 3ʹUTR of CSF1 were obtained by PCR from CSF1 plasmid (Origene, Rockville, MD, USA) and subcloned into the pCMV-NFlag vector (Beyotime Biotechnology, Jiangsu, China).

Techniques: Expressing, Control, In Situ Hybridization, Staining, Immunohistochemical staining