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. Author manuscript; available in PMC: 2014 Jul 25.
Published in final edited form as: Br J Haematol. 2013 Jun 25;162(5):621–630. doi: 10.1111/bjh.12440

Figure 2. LITAF does not induce TNFα secretion upon LPS exposure in B cells.

Figure 2

(A) LITAF expression assessed by Western blot in 9 cell lines of Diffuse Large B Cell Lymphoma (DLBCL) incubated with LPS during 24h, using actin detection as loading control. (B) TNFα secretion measured by ELISA in THP1 and OCI-Ly10 cells treated with LPS for 24 hours, in RL and SC-1 cells harboring a tetracycline inducible LITAF expression plasmid (Tet on LITAF) or the corresponding empty vector (Tet on Empty) in presence of 50 ng/mL doxycycline, and in KARPAS-231 (K231) and VAL cells transfected with a LITAF specific siRNA or non-targeting siRNA (Ctrl.). Subcellular fractionation was performed in four DLBCL cell lines as previously described (Beltran, et al 2011) and LITAF was detected by Western blot (C) β-tubulin and LAMIN A/B were also analyzed as markers of the cytoplasmic “C” and nuclear “N” fractions, respectively. The induction of LITAF expression (D) and the silencing of LITAF (E) were confirmed by Western blot. The effect on cell viability of LITAF over-expression induced by doxycycline exposure in SC-1 (F) and RL (G) cells transfected with a tetracycline inducible expression vector was measured by MTS assays. The results are normalized to the corresponding cells not treated with doxycycline. Error bars represent the standard deviation of three independent experiments.