Figure 5

Muc-1^KO BMDM express very low levels of the chemokine CCL2 and chemokine receptor CCR2. (A) RNA was extracted from WT and Muc-1^KO BMDMs stimulated with LPS (100 ng/ml) for 0, 1, 2, or 4 hours. Quantitative RT-PCR was performed and average gene expression as compared to time 0 is shown for CCL2. * p < 0.05 indicates statistical significance of CCL2 mRNA expression level of WT BMDM as compared to the Muc-1^KO BMDM at 2 hours and 4 hours LPS treatment time points. (B) IHC staining of CCL2 on pancreas sections from WT and Muc-1^KO mice with/without CVB3 infection 8-days post-inoculation showed that CCL2 levels were significantly lower in CVB3 infected Muc-1^KO mice pancreas compared to WT infected mice. (C) BMDM from WT and Muc-1^KO mice were treated with or without LPS (100 ng/ml) for 16 hours and cells were processed for flow cytometric analysis. CD11b+Gr1+ cells were gated and the expression level of CCR2 were compared. Flow cytometric data shown are representative of 3 independent experiments. (D) Quantitative data showed that the median fluorescent intensity (MFI) of CCR2 expression levels were compared between the WT and Muc-1^KO BMDM. Quantification data shown are mean ± SD. **p ≤ 0.01 indicates statistical significance. (E) The mRNA expression level of CCR2 were evaluated by real-time PCR in pancreas from WT and Muc-1^KO mice with CVB3 infection at 4 days, 6 days and 8 days post-inoculation. **p ≤ 0.01, ****p ≤ 0.0001 indicates statistical significance. (F) IHC staining of CCR2 on pancreas sections from WT and Muc-1^KO mice with CVB3 infection 8-days post-inoculation.