Fig. 5

FAM46C is induced in activated B lymphocytes and localizes mainly in the cytosol. a Splenocytes from FAM46C ^{-FLAG +/+} knock-in mice and control animals were isolated and then cultured in presence of IL4 and LPS for 72 h. The level of FAM46C-FLAG was checked by western blot in non-activated (lanes 1–2) and activated (lanes 2–4) cells. GRP94 was used as activation marker, while DBC1 was used as a loading control. b Endogenous FAM46C localizes mainly in the cytosol. Splenocytes from WT and FAM46C ^-FLAG+/+ mice were isolated and fractionated. The whole-cell (T), cytoplasmic (CE), and nuclear (NE) extracts were analyzed by western blot. FAM46C-FLAG was detected using an anti-FLAG antibody. GAPDH and GRP94 were used as a cytosolic markers, while histone H4 and PSPC1 were used as nuclear markers. Arrow indicates the position of FAM46C-FLAG, while asterisks indicts nonspecific bands detected by antibodies, which were present in samples isolated from both WT and FAM46C-FLAG animals