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. 2021 Mar 18;14(3):dmm047589. doi: 10.1242/dmm.047589

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© 2021. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 3. — MCPIP1 suppresses the activation of macrophages and, indirectly, Th17 lymphocytes and B-cell subsets. (A) The total number of CD11c⁺ dendritic cells and F4/80⁺ macrophages and their activation in spleens was quantified by flow cytometry in both investigated genotypes as described in the Materials and Methods. Data represent means±s.e.m. from at least eight mice per group. **P<0.01, ***P<0.001 versus control mice. (B) Spleen sections were stained for CD3⁺ T cells and quantified with image software as described in the Materials and Methods (magnification ×100). Data are means±s.e.m. from at least eight mice per group; ***P<0.001. RNA was isolated from spleens, and mRNA levels in spleen were analyzed for relevant transcripts. (C) Expression levels of Il2 were quantified by real-time PCR. Data are shown as means of the ratio of the specific mRNA versus that of Gapdh mRNA; *P<0.05. (D,E) The total numbers of the CD4⁺ T-cell subset (D) and Th17/Treg ratio (E) were quantified by flow cytometry in both investigated genotypes as described in the Materials and Methods. Data represent means±s.e.m. from at least eight mice per group. *P<0.05 versus control mice. (F) mRNA levels of T-cell-relevant transcription factors in spleens were quantified by real-time PCR. Data are shown as means of the ratio of the specific mRNA versus that of Gapdh mRNA; *P<0.05, ***P<0.001 versus control mice. (G) Spleen sections were stained for CD19⁺ B cells and quantified with image software as described in the Materials and Methods (magnification ×200). Data are means±s.e.m. from at least eight mice per group; ***P<0.001. The total numbers of spleen B220⁺/CD19⁺/MHCII⁺ cells were quantified in 6-month-old mice by flow cytometry. Data are means±s.e.m. of at least eight mice per group; **P<0.01, ***P<0.001 versus control mice. (H) mRNA levels in the spleen were analyzed for B-cell survival and stimulatory factors. Expression levels were quantified by real-time PCR. Data are shown as means of the ratio of the specific mRNA versus that of Gapdh mRNA; *P<0.05, ***P<0.001. (I) Mature B cells and kappa light chain⁺ CD138⁺ (also known as Sdc1⁺) plasma cells were quantified in 6-month-old mice by flow cytometry. Data are means±s.e.m. of at least eight mice per group; **P<0.01 versus control mice. n.s., not significant.