Fig. 5
Over-expression of sOPN and iOPN in OPN-/-Faslpr/lpr DLBCL cell lines and evaluation of TLR9-MYD88 cascade. A. H&E (upper panels) and IF for PAX5/BCL6 (middle panels) performed on OPN-/-Faslpr/lpr mouse spleens from which OPL239 and OPL241 derive. H&E staining on tumorigenic spleens from nu/nu mice intravenously injected with OPL239 and OPL241 (lower panels). Scale bar: 100 μm. B. RT-PCR showing the expression of Bcl6, Bcl2, Irf4 and c-myc in OPL239 and OPL241 cell lines. CD19 + from BALB/c and OPN-/-mice were used as controls. Values are shown as 2^-DCT. C. Western blot on OPL239 and OPL241 lysates illustrating the level of BCL2, c-MYC, IRF4 and the basal level/activation of STAT3 and NF-κB (p65) pathways. D. Western blot for phospho- and total STAT3, phospho- and total p65 (NF-κB) performed on OPL239, OPL239 IRES-Green, OPL239 Spp1-IRES-Green and OPL239 iOPN-IRES-Green with or without 30-min stimulation with CpG 1826. E. RT-PCR for Il6 and Tnfa on OPL239 cell variants with or without 3 h-stimulation with CpG 1826. Values are shown as 2^-DCT. (*, p < 0.05; Two-way ANOVA), (**, p < 0.01; Two-way ANOVA), (***, p < 0.001; Two-way ANOVA). F. Western blot on OPL241 variants illustrating the expression of phospho- and total STAT3 and NF-κB with or without 30-min stimulation with CpG 1826. G. RT-PCR for Il6 and Tnfa on OPL241 cell variants with or without 3 h-stimulation with CpG 1826. Values are shown as 2^-DCT. (*, p < 0.05; Two-way ANOVA), (**, p < 0.01, Two-way ANOVA)