Extended Data Fig. 4: NF-κB-dependent signaling and cytokine secretion from splenocytes and bone marrow derived pDCs is not increased in vikala mice.
From: A TNIP1-driven systemic autoimmune disorder with elevated IgG4
a-d, Immunoblots of splenic B cell lysates from Tnip1+/+ (n = 1) and Tnip1vik/vik (n = 1) mice probed for TNIP1, IκBα and Actin proteins after stimulation with CpG-B, R848 (a), LPS (b), BCR+CpG-B, BCR+R848 (c), BCR (αIgM) and CD40 (d). B cells from 40-week (a, d), 10–12-week (b), and 28–29-week old mice (c). (Densitometric ratio of IκBα to Actin loading control in stimulated wildtype (+/+) and vikala homozygote (vik/vik) B cells shown (right) (a-d). Asterisks indicate poor quality bands that were prohibitive to accurate quantification. e, j, Immunoblots of bone marrow derived pDCs from 16–20 week-old Tnip1+/+ (n = 1) and Tnip1vik/vik (n = 1) mice probed for IκBα (e), TNIP1 (j) and Actin proteins after stimulation with CpG-A and R848. Densitometric ratio of IκBα to Actin loading control shown (e). f-i, Bone marrow derived pDCs from 16–20 week-old Tnip1+/+ (n = 1) and Tnip1vik/vik (n = 1) mice were seeded in 96 well plates and treated with either CpG-A, R848, R837 or infected with E.coli. Supernatants were harvested for quantification by ELISA at 6 h (f-i) post treatment. Bars represent means with standard error of the mean (SEM) and each dot a single mouse (biological replicate). Two-way ANOVA with Sidak’s multiple comparisons; ns, not significant. Data in panels (a-e, j) are representative of experiments performed twice and (f-i) from experiments performed three times.
