Figure 5From: PIAS1 interacts with FLASH and enhances its co-activation of c-MybFLASH, PIAS1 and c-Myb are all co-localized in active RNA polymerase II foci. (A) CV-1 cells were transfected with plasmids encoding HA-PIAS1 and 3×FLAG-FLASH. Cells were analysed by indirect immunofluorescence and confocal microscopy. PIAS1 was detected with rabbit anti-HA antibody and Alexa Fluor 488 goat anti-rabbit IgG (green signal). RNA polymerase II phosphorylated on Ser-5 in the CTD was detected with a mouse monoclonal anti-pol II (8A7) IgM antibody and Alexa Fluor 546 goat anti-mouse IgM, specific for the IgM heavy chains (red signal). FLASH was detected with mouse anti-FLAG antibody and Alexa Fluor 633 goat anti-mouse IgG1, specific for the IgG1 heavy chains (blue signal). Double merged images are shown as indicated, and triple merged images are visualized in right panel (merge). (B) CV-1 cells were transfected with plasmids encoding c-Myb-HA and FLAG-PIAS1. c-Myb was detected with rabbit anti-HA antibody and Alexa Fluor 488 goat anti-rabbit IgG (green signal). Active RNA polymerase II was detected as in (A) (red signal). PIAS1 was detected with mouse anti-FLAG antibody and Alexa Fluor 633 goat anti-mouse IgG1, specific for the IgG1 heavy chains (blue signal). Double merged images are shown as indicated, and triple merged images are visualized in right panel (merge).Back to article page