Journal: Current Biology

Article Title: Gut microbiota suppress feeding induced by palatable foods

doi: 10.1016/j.cub.2022.10.066

Figure Lengend Snippet:

Article Snippet: Rabbit anti-c-Fos 9F6 mAb , Cell Signaling Technologies , Cat#2250; RRID: AB_2247211.

Techniques: Recombinant, DNA Extraction, SYBR Green Assay, Sequencing, Software, Purification

Journal: Molecular Vision

Article Title: Activator protein-1 contributes to the NaCl-induced expression of VEGF and PlGF in RPE cells

doi:

Figure Lengend Snippet: High extracellular NaCl induces elevation in the levels of c-Fos and phosphorylated c-Jun proteins in RPE cells. The cells were stimulated for 2, 4, and 6 h with high (+ 100 mM) NaCl or the hypoxia mimetic CoCl 2 (150 µM). Protein levels were determined with western blot analysis of cell lysates ( A – C ) and nuclear extracts ( D ). A : Example of a western blot analysis in one cell line. Left: The levels of the following proteins were determined: β-actin, c-Fos, phosphorylated c-Fos (P-c-Fos), c-Jun, and phosphorylated c-Jun (P-c-Jun). Right: Negative control obtained with the omission of the first antibodies. B : Cytosolic levels of c-Fos ( left ) and phosphorylated c-Fos proteins ( right ), as determined with densitometric analysis of western blot data. C : Cytosolic levels of c-Jun ( left ) and phosphorylated c-Jun proteins ( right ). The data are normalized to the level of the β-actin protein and are expressed as a percentage of the unstimulated control (100%). Each bar represents data obtained in three to six independent experiments using cell lines from different donors. Significant difference versus unstimulated control: * p<0.05. D : Examples of western blot analysis of the nuclear extracts. The cells were stimulated for 2 and 6 h with high (+ 100 mM) NaCl. Note that the nuclear levels of histone H3 (His H3) and CREB proteins did not change in response to high NaCl.

Article Snippet: The following antibodies were used: a rabbit anti-human β-actin (1:1,000; Cell Signaling, Frankfurt, Germany), a rabbit anti-c-Fos (1:1,000; Cell Signaling), a rabbit anti-c-Jun (1:1,000; Cell Signaling), a rabbit anti-phosphorylated c-Fos (1:1,000; ThermoFisher Scientific, Waltham, MA), a rabbit anti-phosphorylated c-Jun (1:1,000; Cell Signaling), a rabbit anti-cAMP response element-binding protein (CREB; 1:1,000; Cell Signaling), a rabbit anti-histone H3 (1:1,000; Cell Signaling), and anti-rabbit immunoglobulin G (IgG) conjugated with alkaline phosphatase (1:2,000; Cell Signaling).

Techniques: Western Blot, Negative Control