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. 2013 Jun 3;10(4):360–372. doi: 10.1038/cmi.2013.14

Figure 4.

Figure 4

CpG ODN-induced apoptosis of CH27 lymphoma B cells depends on Fas and Fas ligand. (A) splenic B cells (a and d) and TLR9 expressing CH27 cells (b–c and e–h) were treated with 10 µg/ml GpC or CpG ODNs for 48 h and stained for CD95 (Fas) or CD178 (Fas ligand). The surface levels of CD95 (a–c) and CD178 (d–f), as well as cell sizes (g–h) were analyzed by flow cytometry. A population of CpG-treated CH27 cells with reduced sizes (R2) were gated, and their CD95 and CD178 expression levels were measured (c and f). Representative flow cytometry plots are shown. (B) The mean (±s.d.) fold increases of the mean fluorescence intensity of CD95 and CD178 in CpG-treated cells compared to GpC-treated cells from three independent experiments are shown. (C) CH27 clone 1 cells were incubated with 1 or 10 µg/ml GpC or CpG ODN in the presence of an isotype control antibody (Iso) or Fas ligand-neutralizing antibody (anti-Fas, 10 µg/ml) for 48 h. The percentages of Ann V+ PI− cells were determined. The mean (±s.d.) of three independent experiments are shown. *P<0.01. AnnV, Annexin V; CpG, cytosyl guanosyl; ODN, oligodeoxynucleotide; PI, propidium iodide; TLR, Toll-like receptor.