Fig. 5: LSD1/CoREST complex physically interacts with TCF1 and antagonizes its transcriptional activity.
From: LSD1 inhibition sustains T cell invigoration with a durable response to PD-1 blockade
a MFI of TCF1 in CD8+TCF1+PD-1int TILs analyzed by flow cytometry (n = 4 per group). b The physical interaction between LSD1 and TCF1 isoforms as well as other proteins was examined by co-immunoprecipitation (co-IP) assays using whole-cell extract (WCE) of HEK293T cells transfected with FLAG-HA-tagged LSD1. c Schematic representation of different isoforms of mouse TCF1. d Co-IP assays precipitating HA-tagged TCF1 isoforms and detecting LSD1 in eluted IP fractions. e Co-IP assays precipitating Flag-tagged TCF1 (referring to p45 unless otherwise specified) and detecting components of the LSD1/CoREST complex in eluted IP fractions. f–j TCF/LEF reporter assay measuring relative luciferase units (RLU) in response to transfection with plasmids expressing indicated proteins (f–j, n = 4 per condition) or shRNA targeting LSD1 (i) and the treatment with iCRT3 (i) (n = 4 per condition). Protein expression for plasmid transfection in f is shown by immunoblotting (g). k, l Representative flow plots (k) and Slamf6 MFI (l) of splenic CD8+ T cells activated in vitro for 3 days with anti-CD3/anti-CD28 in the presence of vehicle, GSK2879552 or iCRT3 (n = 3 per treatment). Data represent two (a, b, d–j) or three (k, l) independent experiments and are presented as mean ± SEM (a) or mean ± SD (f, h–j, l). Statistical significance was determined by two-sided unpaired t test (a, f, h–j, l). Source data are provided as a Source data file.
