Fig. 2
Evaluation of STAT3 pathway activation in CD19 + B cells from autoimmune mice. A. Western blot illustrating the expression of phospho- and total STAT3 in splenic CD19 + B cells from Faslpr/lpr (n = 3) and OPN-/-Faslpr/lpr (n = 3) mice. CD19 + from a BALB/c and a OPN-/- mouse were used as controls. B. Quantification of western blot, relative to β-ACTIN as housekeeping gene of ph-STAT3 (Student t test; ***, p:0.0007) and total STAT3 (ns, p: 0.1237). Statistics was applied to values related to Faslpr/lpr and OPN-/-Faslpr/lpr B cells. C. Representative IHC (upper panels) showing the expression of ph-STAT3 in OPN-sufficient and -deficient lymphomas. Scale bar: 100 μm. Representative IF (lower panels) for PAX5 (pan B cell marker) and ph-STAT3 on OPN + / + and -/-tumours. Scale bar: 50 μm. D. RT-PCR on purified splenic CD19 + cells from Faslpr/lpr and OPN-/-Faslpr/lpr mice showing the expression of STAT3- target genes Prdm1 (Student t test; *, p: 0.0261) and Birc5 (Student t test; *, p: 0.0195). CD19 + from B/c and OPN-/- mice were used as controls. Values are shown as 2^-DCT. Data are a pool of two independent experiments with 3 samples for each group. E. Representative RNA scope imaging showing Prdm1 expression on spleens from autoimmune mice