Fig. 2.

C1q gene expression is decreased in Mafb ^−/− macrophages. a Messenger RNAs of C1q genes in PMs were examined by qRT-PCR (n = 3 for each group). b PMs of Mafb ^f/f::LysM-Cre and Mafb ^f/f were analyzed for the expression of the Mafb and C1q genes (n = 4 for each group). c A western blot analysis was performed to measure C1q protein levels in the PMs of WT and Mafb ^−/−. The protein extracts were normalized to β-actin expression. The data are from one experiment that was representative of at least two independent experiments. d The C1q genes were analyzed by qRT-PCR of macrophages and immature DCs from WT or Mafb ^−/− (n = 4 for each group). The data are from one experiment that was representative of at least two independent experiments. a, b, d The data were normalized to Hprt mRNA and are presented as the mean ± s.e.m. *p < 0.05 compared with WT, **p < 0.01 (Student’s t-test). e, f The hemolysis rates were analyzed using serum from fetal liver cell-transplanted mice 3 months after transplantation (e) or Mafb ^f/f::LysM-Cre and control mice (8–10 weeks old) (f). The data were analyzed with the Mann–Whitney U-test, and the results from two independent experiments were pooled. g A western blot analysis was performed to measure the C1q protein levels in the serum of 8-week-old WT and Mafb ^−/− mice. Lower panel, the data were normalized to the protein band of Ponceau S and are presented as the mean ± s.e.m. **p < 0.01 compared with Mafb ^f/f (Student’s t-test). The data are from one experiment that was representative of at least two independent experiments. h THP-1 cells were transfected with either control siRNA (si-control) or MAFB siRNA (si-MAFB). The cells were stimulated with dexamethasone and IFNγ for 24 h after PMA stimulation. The data were normalized to HPRT mRNA and are presented as the mean ± s.e.m. n = 4 for each group. *p < 0.05 compared with si-control (Mafb, C1qa, C1qc, Student’s t-test; C1qb, Welch’s t-test). The data are from one experiment that was representative of two independent experiments. i qRT-PCR was conducted using heterozygous mafb ^b337/+ mutant zebrafish. The data were normalized to csf1r mRNA and are presented as the mean ± s.e.m. WT; n = 7, mafb ^b337/+; n = 6 for each group. *p < 0.05 compared with WT (Student’s t-test). The data are from one experiment that was representative of two independent experiments