Inducible expression of immediate early genes is regulated through dynamic chromatin association by NF45/ILF2 and NF90/NF110/ILF3
Fig 7
Reduced NF90/NF110 expression attenuated inducible expression IEGs by immunofluorescence analysis.
HEK293 cells stably transfected with pINDUCER10-shNF90/NF110 (D2) or pINDUCER10-shNF45 (D5) were treated without or with doxycycline for 96 h, then seeded on chamber slides, and serum starved for 24 h and stimulated by PMA (20 ng/ml) for 2 h for maximal protein expression of IEG. Slides were incubated with mouse monoclonal antibodies against NF90/NF110, and rabbit monoclonal antibodies against EGR1 (a-b), FOS (c-d), or JUN (e-f), then incubated with fluorescent conjugated secondary antibodies anti-mouse Alexa Fluor 488 and anti-rabbit Alexa Fluor 594. Slides were counterstained with DAPI, then visualized by confocal fluorescent microscopy. Panels (b,d,f) show doxycycline-mediated shRNA knockdown compared to (a,c,e) respectively.