Depletion of THOC5/FMIP causes down-regulation of THOC1. (A): Liver extracts from three times poly (I:C) 250 μg treated (+) (14 days) and untreated (-) six-week-old Mx-cre THOC5/FMIP (flox/flox) mice (n = 3) were examined by THOC1, Aly, THOC6, THOC5/FMIP, Actin and Histone H3 specific immunoblotting. Each sample consisted of 100 μg of liver. (B): Liver or bone marrow extracts from poly (I:C) treated (+) (14 days) were examined by THOC1, THOC5/FMIP, and actin specific immunoblotting. THOC1*: 60 kDa THOC1 degradation product from poly (I:C) treated liver or bone marrow sample. (C): mRNA was isolated from the same liver samples and THOC5/FMIP (643 bp and 431 bp (without exons IV/V)), THOC1 (258 bp), Aly (420 bp) and Actin (509 bp) specific RT-PCR were performed. (D): MEF THOC5/FMIP (flox/flox) cells were infected with adenovirus carrying GFP gene (Ad-GFP) or adenovirus carrying GFP and cre-recombinase genes (Ad-GFP-Cre) (M.O.I. 50; Vector Biolab, Philadelphia, PA, USA) then lysed by laemmli buffer. Extracts were then examined by THOC1, Aly, THOC6, THOC5/FMIP, GAPDH and Histone H3 specific immunoblotting.
Mancini et al. BMC Biology 2010 8:1 doi:10.1186/1741-7007-8-1