Journal: Cells

Article Title: IL-1β and TNFα Cooperativity in Regulating IL-6 Expression in Adipocytes Depends on CREB Binding and H3K14 Acetylation

doi: 10.3390/cells10113228

Figure Lengend Snippet: Combined effect of IL-1β and TNFα on IL-6 expression in mouse adipocytes. ( A , B ) 3T3 L preadipocytes were differentiated into adipocytes as described in materials & methods. Lipid droplets in adipocytes were determined by using Nile Red staining. Morphology of adipocytes and adipogenic markers were shown. Scale Bar 50 µm. Mouse 3T3-L adipocytes were stimulated with IL-1β (10 ng/mL) and TNFα (10 ng/mL) alone or in combination for 24 h. Cells and culture media were collected. ( C ) Total RNA was extracted from the cells and IL-6 mRNA was quantified by real time PCR. Relative mRNA expression was expressed as a fold change. ( D ) Secreted IL-6 protein in culture media was determined by ELISA. ( E ) Different number of cells (1, 0.5, 0.25 million) were treated with IL-1β (10 ng/mL) and TNFα (10 ng/mL) alone or in combination for 24 h. Cells and culture media were collected, Secreted IL-6 protein in culture media was determined by ELISA. ( F ) 3T3 adipocyte cells were stained for confocal microscopy, as described in the Materials and Methods section. IL-6 expression is shown by green fluorescence (inset), whereas nuclei are stained blue with DAPI (original magnification ×40). Scale Bar 20 µm. ( G ) IL-6 fluorescence intensity was determined for 10 random images. The results obtained from three independent experiments are shown. All data are expressed as mean ± SEM ( n = 3). ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human preadipocytes derived from subcutaneous and omental visceral adipose tissues from lean and obese individuals were obtained from ZenBio (Research Triangle Park, NC, USA; catalogue numbers: SP-F-1, OP-F-1, and OP-F-3, respectively).

Techniques: Expressing, Staining, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Confocal Microscopy, Fluorescence

Journal: Cancers

Article Title: Adipose Tissue-Derived Mesenchymal Stromal/Stem Cells, Obesity and the Tumor Microenvironment of Breast Cancer

doi: 10.3390/cancers14163908

Figure Lengend Snippet: Functional alterations of breast cancer cells induced by ASCs/MSCs.

Article Snippet: Human primary subcutaneous pre-adipocytes (pre-hASCs, Lonza) , MCF7, T47D, ZR-75-1, SK-BR-3 BC cell lines and murine 3T3-L1 pre-adipocytes in vitro , Conditioned medium of ASCs stimulated proliferation and migration of MCF7, T47D, SK-BR-3, and ZR-75-1 cells. Additionally, supernatant of ASCs upregulated the expression of S100A7 and its knockdown abrogated the tumorigenic effect of ASCs on the tested breast cancer cells. , [ ] .

Techniques: Functional Assay, Derivative Assay, In Vitro, Migration, Activation Assay, In Vivo, Gene Expression, Isolation, Expressing, Knockdown, Transplantation Assay

Journal: Cancers

Article Title: Adipose Tissue-Derived Mesenchymal Stromal/Stem Cells, Obesity and the Tumor Microenvironment of Breast Cancer

doi: 10.3390/cancers14163908

Figure Lengend Snippet: Subtypes of cancer associated ASCs/CAFs in breast cancer.

Article Snippet: Human primary subcutaneous pre-adipocytes (pre-hASCs, Lonza) , MCF7, T47D, ZR-75-1, SK-BR-3 BC cell lines and murine 3T3-L1 pre-adipocytes in vitro , Conditioned medium of ASCs stimulated proliferation and migration of MCF7, T47D, SK-BR-3, and ZR-75-1 cells. Additionally, supernatant of ASCs upregulated the expression of S100A7 and its knockdown abrogated the tumorigenic effect of ASCs on the tested breast cancer cells. , [ ] .

Techniques: Isolation, In Vitro, Inhibition, Gene Expression, Expressing, Migration, Marker, RNA Sequencing, Derivative Assay