Figure 6

TLR7 stimulation modulates p38 and JNK phosphorylation in CLL but has significantly less pronounced or no effect on stereotyped subset #4 cases. (A) Immunoblotting studies revealed that stimulation through TLR7 with imiquimod affects JNK and p38 phosphorylation in various CLL subgroups, both unmutated and mutated, with the significant exception of stereotyped subset #4 cases that are generally also nonresponsive to TLR7 stimulation in terms of costimulatory molecule or apoptosis induction. In this figure seven representative cases are represented. Densitometric analysis of (B) p-JNK(p54)/β-actin (C) p-JNK(p46)/β-actin and (C) p-p38/β-actin levels in the 12 CLL samples analyzed. The graphs represent the median values for each group which determined as the ratio of the optical density (OD) of p-protein and β-actin using the ImageJ software (National Institutes of Health, Bethesda, MD, USA; https://doi.org/imagej.nih.gov/ij/, 1997–2012). IIM, imiquimod; M: mutated; UM: unmutated; non #4 (4_34), cases expressing the IGHV4-34 gene in non-stereotyped rearrangements; #1, stereotyped subset #1; +, positive control (CLL cells treated for 1 h with CpG).