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. 2020 Sep 30;18(4):1016–1031. doi: 10.1038/s41423-020-00552-0

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© CSI and USTC 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — Freshly purified B cells, LPS-stimulated plasmablasts, and plasma cells from B cell-specific STING^KO mice exhibited significantly higher levels of surface BCR and were more responsive to BCR activation than those from STING-proficient mice; TNP-Ficoll-immunized B cell-specific STING^KO mice generated significantly more TNP-specific plasma cells and antibodies than TNP-Ficoll-immunized STING^WT mice. a Freshly purified B cells from CD19Cre (n = 3) and B cell-specific STING^KO (CD19Cre/STING^flox/flox; n = 3) mice were stimulated with LPS (20 μg/mL) for 3 days. Each day, cells were surface stained with B220-BV605 and IgM-PE-Cy7. Gated B220+ populations were analyzed for the expression of IgM. The mean fluorescence intensity (MFI) of IgM was plotted as the mean ± SEM. b CD19Cre (n = 4) and B cell-specific STING^KO (n = 4) mice were intraperitoneally immunized with TNP-Ficoll on Day 0. On Day 9, bone marrow cells from immunized mice were stained with B220-Alexa 488, CD138-PE, and Igβ-APC. Gated B220–/CD138+ plasma cells were analyzed for the expression of Igβ. The MFI of Igβ was plotted as the mean ± SEM. c Freshly purified B cells from STING^WT and STING^KO mice and (d) those B cells stimulated with LPS for 3 days were activated with goat anti-mouse IgM F(ab’)2 (20 μg/mL) for the indicated times and lysed for immunoblot analysis. e-j STING^WT and B cell-specific STING^KO mice were intraperitoneally immunized with TNP-Ficoll on Day 0. Immunized mice were sacrificed on Day 9, and the presence of TNP-specific plasma cells in the spleens and bone marrow was analyzed. Anti-TNP IgM-secreting plasma cells in the spleens (e) and bone marrow (f) of TNP-Ficoll-immunized STING^WT (n = 9) and B cell-specific STING^KO (n = 9) mice were quantified by ELISPOT. Anti-TNP IgG-secreting plasma cells in the spleens (g) and bone marrow (h) of TNP-Ficoll-immunized STING^WT (n = 9) and B cell-specific STING^KO (n = 9) mice were quantified by ELISPOT. Nine days after the single immunization, serum levels of anti-TNP IgM (i) and IgG3 (j) in TNP-Ficoll-immunized STING^WT (n = 11) and B cell-specific STING^KO (n = 11) mice were determined by ELISA