Dysregulated NF-κB activation in Il1r8+/+/lpr and Il1r8−/−/lpr mice.

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Dysregulated NF-κB activation in Il1r8+/+/lpr and Il1r8−/−/lpr mice. Dysregulated NF-κB activation in Il1r8+/+/lpr and Il1r8−/−/lpr mice. A, Western blot of spleen B cells with the indicated antibodies. β-Actin expression was analyzed as internal control. B and C, Densitometric signal ratios of p52/p100 and phospho-p65/p65 shown in A (Mann–Whitney test). n = 4 Il1r8+/+, n = 5 Il1r8−/−, n = 4 Il1r8+/+/lpr, n = 6 Il1r8−/−/lpr mice. D, Real-time PCR array of NF-κB signaling target genes. Expression data are shown only for the genes for which a fold difference (FD) > 2 was observed in at least one comparison between two groups of mice (see Supplementary Table S2 for individual data). In the graph, a two-color scale formatting scheme was used to format cells: red is the maximum expression; blue, minimum. Each column represents one sample (from one mouse). E, Nfkbiz mRNA expression in purified B cells (unpaired Student t test with Welch correction). A–E, One experiment performed. F, FACS analysis of CD86 expression in overnight-cultured purified B cells in basal condition and after LPS stimulation. Top, representative histograms. Bottom, results are reported as MFI normalized on fluorescence minus one. One representative experiment with B cells collected from 3 to 6 mice (1 or 2 replicates per mouse) out of 2 performed is shown (unpaired Student t test with Welch correction). B, C, E, F, mean and SD are shown. Federica Riva et al. Cancer Immunol Res 2019;7:874-885 ©2019 by American Association for Cancer Research