Journal: iScience

Article Title: An appeasing pheromone ameliorates fear responses in the brown rat ( Rattus norvegicus )

doi: 10.1016/j.isci.2023.107081

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti- c -Fos , Cell Signaling Technology , Cat#2250; RRID: AB_2247211.

Techniques: Plasmid Preparation, Recombinant

Journal: Frontiers in Oncology

Article Title: Virtual screening of ultra-large chemical libraries identifies cell-permeable small-molecule inhibitors of a “non-druggable” target, STAT3 N-terminal domain

doi: 10.3389/fonc.2023.1144153

Figure Lengend Snippet: Compounds interfere with STAT3 N-domain signaling. (A) , STAT3 ND binders inhibit STAT3 signaling in HEK-BLUE IL-10 reporter cells. Cells (~70,000 cells/mL) were incubated with IL-10 (0.1 ng/ml) without or with 500μM of compound by measuring secreted embryonic alkaline phosphatase (SEAP) levels induced by 24 hours incubation). (B) . Inhibition of IL-10 induced STAT3 signaling in HEK-BLUE IL-10 reporter cells was concentration-dependent. As expected, compounds exhibited cell toxicity (blue line). However, inhibition of STAT3 activity was more significant and did not appear to be toxicity mediated. (C) , Compounds induce expression of STAT3 ND inhibition marker c-Fos. DU145 cells were treated with compounds for 3 hours (with DMSO treatment as a negative control), and nuclear fraction was isolated and used for Western blot analysis as described. Anti-c-Fos antibody was used at 1:1000 dilution. The blot was stripped and re-probed with β-Actin antibody at 1:1000 to demonstrate equal loading of the wells. (D) . Compounds are selectively toxic to prostate tumor cells (DU145) and show very little toxicity to normal prostate epithelial cells (RWPE-1). (E) , Lead compounds ST3ND-25 and ST3ND-34 are stable in human plasma, but undergo rapid metabolism in mouse plasma.

Article Snippet: C-Fos rabbit monoclonal antibody (9F6, Cell Signaling Technology, Danvers, MA) was diluted 1:1000.

Techniques: Incubation, Inhibition, Concentration Assay, Activity Assay, Expressing, Marker, Negative Control, Isolation, Western Blot