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. 2000 May 15;191(10):1735–1744. doi: 10.1084/jem.191.10.1735

Figure 2.

Figure 2

Anti-IgM induces normal c-rel transcript levels, and amounts of c-rel in xid B cells are normal. (A) Wild-type and xid B splenic cells were cultured for 4 h in medium, F(ab′)2 anti-IgM (10 μg/ml), or CD40 ligand (1:2 dilution), and whole cell extracts were prepared. C-rel Western blot analysis was performed using extracts prepared from 5 × 105 cells. β-actin served as a loading control. (B) Wild-type and xid splenic B cells were cultured for 4 h in medium, F(ab′)2 anti-IgM (10 μg/ml), or PMA and ionomycin (30 nM and 1 μM, respectively), and total RNA was prepared. RT-PCR (top panel) analysis was conducted using primers specific for c-rel (or GαS, which served as a loading control). RNA samples from WEHI231 and T220 cells served as positive and negative controls, respectively, for this assay. Northern analysis (bottom panel) was conducted using 10 μg of each sample. Equal loading was determined by ethidium bromide staining of 18S and 28S ribosomal bands (data not shown). These figures represent one of three experiments.