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. Author manuscript; available in PMC: 2018 Mar 15.
Published in final edited form as: J Immunol. 2017 Feb 10;198(6):2244–2248. doi: 10.4049/jimmunol.1502266

FIGURE 1.

FIGURE 1

Enforced Sox2 expression alters CSR and AID levels. (A) Immunoblot of protein extracts from wild type splenic B cells stimulated with anti-CD40 and IL-4 for the indicated time and probed for expression of Sox2, AID or Gapdh (loading control) (n = 4). (B) Representative immunoblot of purified IgM+ or IgG1+ splenic B cells obtained from anti-CD40 and IL-4 cultures; tubulin, loading control (n=3). (C) Representative immunoblot of activated splenic B cells from wild type or AID−/− mice (n=3). (D) Representative flow cytometry of B cells infected with pMIR-Sox2 or pMIR control vector. pMIR expresses mCherry from an IRES element downstream of the test gene. Cells were gated first on the mCherry+ population as a marker for infection and then on B220+ IgG1+ cells. Bar graph shows CSR of 15 independent experiments; CSR in the pMIR control population was set at 100%. (E) Immunoblot shows expression of Sox2, AID and tubulin (loading control). (F) mCherry+ B cells were sorted 72h after infection and analyzed for Aicda expression by qPCR; Aicda level in pMIR sample was set to 1 (n = 3).