Journal: Febs Letters

Article Title: EWS‐FLI1 impairs aryl hydrocarbon receptor activation by blocking tryptophan breakdown via the kynurenine pathway

doi: 10.1002/1873-3468.12243

Figure Lengend Snippet: EWS ‐ FLI 1 knockdown initiates TRP breakdown and leads to the accumulation of metabolites of the KYN pathway. A673sh cells were grown (EWS‐FLI1‐high, EFH ) or treated with dox ( EWS ‐ FLI 1‐low, EFL ) for 48 or 72 h and cell culture supernatants were analyzed. Prior experiment start, fresh full medium + 10% FCS was added and kept on the cells until collection. Results were normalized to control medium without cells and data are shown as relative fold change over control conditions for EFH cells (white bars) and EFL cells (black bars). (A) TRP decreased in cells where EWS ‐ FLI 1 has been silenced for 72 h, whereas the downstream metabolites of TRP , KYN , and KYNA , were elevated (B, C). (D) 3‐ HK increased slightly faster in the supernatant of EFL than of EFH cells at 48 h of incubation, while no difference in 3‐ HK accumulation was detected after 72 h. Data are shown as means ± SD from four independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Antibodies were: AHR (Cell Signaling Technology, New England Biolabs GmbH, Frankfurt, Germany), FLI1 (MyBiosource, San Diego, CA, USA), Lamin A/C (Santa Cruz, Dallas, TX, USA), TDO2 (Abcam, Cambridge, UK), Vinculin (Sigma, Darmstadt, Germany), α‐Tubulin (Calbiochem, San Diego, CA, USA).

Techniques: Knockdown, Cell Culture, Control, Incubation