Journal: International Journal of Molecular Sciences

Article Title: Tumor Promoting Effect of BMP Signaling in Endometrial Cancer

doi: 10.3390/ijms22157882

Figure Lengend Snippet: BMP2 promotes EC cell stemness by c-KIT induction. ( A ) BMP2 stimulation induces SMAD1/5/8 phosphorylation in Ishikawa cells. Cells were treated in the absence (CT) or presence of 20 ng/mL BMP2 and 200 nM LDN193189 (LDN) for 24 h. α-tubulin was used as an internal control. ( B ) BMP2 induces stemness of Ishikawa cells, as determined by a sphere formation assay. Cells were cultured with stem cell medium containing 20 ng/mL BMP2 and 200 nM LDN in 96-well ultra-low attachment plates for eight days; thereafter, sphere numbers per well were counted using a microscope. Images of spheres are shown at the bottom of the graphs. Scale bar = 200 µm. ( C ) BMP2 induces expression of CD44 and c-KIT mRNA in Ishikawa cells. Cells were treated with PBS (CT), BMP2 (20 ng/mL) or LDN, alone or in combination, for 72 h. mRNA expression was determined by RT-PCR and is shown as fold change relative to control (CT). ( D ) c-KIT expression was quantified by immunoblots in Ishikawa cells 72 h after transfection with empty vector (CT) or c-KIT (KIT) plasmids. α-tubulin was used as an internal control. ( E ) Overexpression of c-KIT by transfection induces stemness of Ishikawa cells, as determined by a sphere formation assay. ( F , G ) BMP2-induced stemness of Ishikawa cells is dependent on c-Kit. Cells were transfected with siNC, siKIT-1, or siKIT-2 for 48 h; thereafter, cells were cultured for an additional eight days in the presence and absence of 20 ng/mL BMP2 ( F ), or incubated in the presence and absence of 20 ng/mL BMP2 and 10 µM imatinib ( G ). Cancer stemness was determined by the formation of spheres. ( H , I ) Ishikawa cells were incubated in the absence (CT) or presence of 200 nM LDN and 500 µM carboplatin (CBDCA). Cell proliferation was determined after 72 h by an MTS assay, and is expressed relative to CT ( H ), and stemness by a sphere formation assay after eight days ( I ). The results in panels B, C, E–I are shown as the mean ± SE. * p -value < 0.05, ** p -value < 0.01.

Article Snippet: Negative controls (siNC) were from the Stealth RNAi siRNA Negative Control Kit (Invitrogen). c-KIT expression plasmid was purchased from Origene (SC120061; Rockville, MD, USA) and transfected into Ishikawa cells using Lipofectamine 2000 transfection reagent (Invitrogen). pcDNA 3.0 (Invitrogen) was used as a control (CT).

Techniques: Tube Formation Assay, Cell Culture, Microscopy, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Plasmid Preparation, Over Expression, Incubation, MTS Assay

Journal: Microorganisms

Article Title: Framework as a Service, FaaS: Personalized Prebiotic Development for Infants with the Elements of Time and Parametric Modelling of In Vitro Fermentation

doi: 10.3390/microorganisms8050623

Figure Lengend Snippet: Heatmap showing short-chain fatty acid production characteristics, dissolved ammonia production characteristics, and total probiotic population pattern characteristics during the 205-h fermentation of carbohydrates by infant fecal inoculum. ( a ) Maximum SCFA production rates; ( b ) increase of total SCFA concentration; ( c ) lag time; ( d ) production characteristics of dissolved ammonia; ( e ) population increase characteristics of total probiotic population pattern; ( f ) population decline characteristics of total probiotic population pattern. Green color stands for high while pink color stands for low. Commensal bifidobacteria are enumerated as total probiotics.

Article Snippet: External standards include acetic, propionic, butyric, valeric, and caproic acids (SCFA standards kit, Alltech Inc., Nicholasville, Kentucky, USA).

Techniques: Concentration Assay, Probiotics