serum free sf dmem  (MedChemExpress)

Journal: PLOS ONE

Article Title: A xenotransplantation mouse model to study physiology of the mammary gland from large mammals

doi: 10.1371/journal.pone.0298390

Figure Lengend Snippet: (a). Sirius red and Masson’s Trichrome (MTC) histological stains, and IHC analysis of vimentin presence in vehicle-injected and mammary fibroblast pre-implanted MFPs (2 weeks post-injection). Scale bar = 200 μm. (b). Acetocarmine-stained mammary xenografts from vehicle-injected and mammary fibroblast pre-implanted MFPs. Black dotted lines designate xenograft borders. Arrowheads indicate regions of xenograft blebbing. Scale bar = 500 μm. (c). Surface area (mm 2 ) quantifications of acetocarmine stained mammary xenografts within vehicle-injected or fibroblast pre-implanted MFPs. Error bars represent standard deviation (SD). Statistical significance was assessed using unpaired (Student’s) t-test (P ≤ 0.05). (d). BrdU assay assessing equine and canine MDEC proliferation over 24 h treatment with species-matched mammary fibroblast CM compared to control base media (DMEM). Symbols (circle, square, and triangle) represent individual MDEC cultures of both species. Error bars represent standard deviation (SD). Statistical significance was assessed using paired two-tailed t-tests (P ≤ 0.05).

Article Snippet: Four ml of serum-free (sf) DMEM (Corning Inc.) was added to fibroblast cell monolayers and incubated for 24 h to generate fibroblast CM.

Techniques: Injection, Staining, Standard Deviation, BrdU Staining, Control, Two Tailed Test

Journal: Chinese Medical Journal

Article Title: Vimentin modulates apoptosis and inflammatory cytokine release by a human monocytic cell line (THP-1) in response to lipopolysaccharides in vitro

doi: 10.1097/CM9.0000000000000187

Figure Lengend Snippet: Effect of vimentin suppression by siRNA on cell survival assessed by TUNEL assay. THP-1 cells were transfected with vimentin-specific or scramble siRNA followed by exposure to LPS (10 μg/mL), and apoptosis was assessed by TUNEL assay as described in the methods. (A–D) Representative images of TUNEL assay. Green: positive TUNEL staining; blue: nuclei staining with DAPI. (E) Average of three separate TUNEL assay results. Vertical axis: TUNEL positivity (%); horizontal axis: cells transfected with control or vimentin-specific siRNA. Open bar: cells cultured in SF-DMEM only; closed bar: cells treated with LPS. ∗ P < 0.05. LPS: Lipopolysaccharide; SF-DMEM: Serum-free Dulbecco modified Eagle media; TUNEL: TdT-mediated dUTP Nick-End Labeling.

Article Snippet: For siRNA transfection, cells were washed once with serum-free DMEM (SF-DMEM; Life Technology) without antibiotics and amphotericin B followed by plating in 60 mm dishes (3 × 10 6 cells/dish) in 1 mL Optimal-Minimal Essential Medium (Opti-MEM; Life Technology).

Techniques: TUNEL Assay, Transfection, Staining, Cell Culture, Modification, End Labeling

Journal: Chinese Medical Journal

Article Title: Vimentin modulates apoptosis and inflammatory cytokine release by a human monocytic cell line (THP-1) in response to lipopolysaccharides in vitro

doi: 10.1097/CM9.0000000000000187

Figure Lengend Snippet: Effect of vimentin over-expression on cell survival assessed by TUNEL assay. THP-1 cells were transfected with a plasmid containing either a negative control vector or vimentin gene followed by exposure to LPS (10 μg/mL), and apoptosis was assessed by TUNEL assay as described in the methods. (A–D) Representative images of TUNEL assay. Green: positive TUNEL staining; blue: nuclei staining with DAPI. (E) average of three separate TUNEL assay results. Vertical axis: TUNEL positivity (%); horizontal axis: cells transfected with control or vimentin-specific siRNA. Open bar: cells cultured in SF-DMEM only; closed bar: cells treated with LPS. ∗ P < 0.05. LPS: Lipopolysaccharide; SF-DMEM: Serum-free Dulbecco modified Eagle media; TUNEL: TdT-mediated dUTP Nick-End Labeling.

Article Snippet: For siRNA transfection, cells were washed once with serum-free DMEM (SF-DMEM; Life Technology) without antibiotics and amphotericin B followed by plating in 60 mm dishes (3 × 10 6 cells/dish) in 1 mL Optimal-Minimal Essential Medium (Opti-MEM; Life Technology).

Techniques: Over Expression, TUNEL Assay, Transfection, Plasmid Preparation, Negative Control, Staining, Cell Culture, Modification, End Labeling

Journal: Chinese Medical Journal

Article Title: Vimentin modulates apoptosis and inflammatory cytokine release by a human monocytic cell line (THP-1) in response to lipopolysaccharides in vitro

doi: 10.1097/CM9.0000000000000187

Figure Lengend Snippet: Effect of vimentin suppression on caspase-3 and Bcl-2. THP-1 cells were transfected with vimentin-specific or scramble siRNA followed by exposure to LPS (10 μg/mL), and caspase-3 and Bcl-2 were assessed by immunoblotting as described in the methods. (A) Representative images of immunoblotting. (B, C) Average of three separate immunoblottings for caspase-3 (B) and Bcl-2 (C). Vertical axis: ratio to control; horizontal axis: cells transfected with control or vimentin-specific siRNA. Open bar: cells cultured in SF-DMEM only; closed bar: cells treated with LPS. ∗ P < 0.05. LPS: Lipopolysaccharide; SF-DMEM: Serum-free Dulbecco modified Eagle media.

Article Snippet: For siRNA transfection, cells were washed once with serum-free DMEM (SF-DMEM; Life Technology) without antibiotics and amphotericin B followed by plating in 60 mm dishes (3 × 10 6 cells/dish) in 1 mL Optimal-Minimal Essential Medium (Opti-MEM; Life Technology).

Techniques: Transfection, Western Blot, Cell Culture, Modification

Journal: Chinese Medical Journal

Article Title: Vimentin modulates apoptosis and inflammatory cytokine release by a human monocytic cell line (THP-1) in response to lipopolysaccharides in vitro

doi: 10.1097/CM9.0000000000000187

Figure Lengend Snippet: Effect of vimentin suppression or over-expression on cytokine release. Vimentin was either suppressed by siRNA or over-expressed by transfecting a plasmid containing vimentin promoter followed by exposure to LPS (10 μg/mL), and cytokines were quantified by ELISA as described in the methods. (A) IL-6. (B) TNF-α. (C) IL-10. Vertical axis: cytokine concentration (pg/day/10 5 cells); horizontal axis: cells transfected with control-siRNA, vimentin-specific siRNA or a plasmid containing vimentin promoter. Open bar: cells cultured in SF-DMEM only; closed bar: cells treated with LPS. Data presented were an average of three separate experiments. ∗ P < 0.05. IL: Interleukin; LPS: Lipopolysaccharide; SF-DMEM: Serum-free Dulbecco modified Eagle media.

Article Snippet: For siRNA transfection, cells were washed once with serum-free DMEM (SF-DMEM; Life Technology) without antibiotics and amphotericin B followed by plating in 60 mm dishes (3 × 10 6 cells/dish) in 1 mL Optimal-Minimal Essential Medium (Opti-MEM; Life Technology).

Techniques: Over Expression, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Transfection, Cell Culture, Modification