Characterization of coilin phosphomutant localization in endogenous coilin reduced stable cell lines.

<p>GFP-coilin and GFP-coilin phosphomutant stable cell lines were transfected with control (Ctrl) or coilin siRNA (KD) for 24 h. 24 h of post siRNA treatment, cells were treated with doxycycline and incubated for another 24 h. The 48 h siRNA transfected and 24 h doxycycline induced stable cell lines were fixed, extracted and immunostained for SMN (red). Nuclei were stained with DAPI (blue). Arrows mark some canonical CBs (containing coilin and SMN). Double arrows mark SMN foci that lack coilin (Gems). Single arrowheads indicate nucleolar coilin accumulation. Double arrowheads mark coilin foci that do not have significant enrichment of SMN, and triple arrowheads indicate dim micro-CB structures. Note that the GFP-coilin ON signal was difficult to detect after the siRNA transfection protocol, so polyclonal GFP antibodies were used to amplify this signal. Scale bars 10 µm.