M2-mCherry reporter construct recapitulates IL10-dependent expansion of M2-transduced primary B cells in vitro.

<p>(A) Retroviruses were derived from replication defective murine stem cell virus (MSCV) vectors that drive constitutive expression of the wild type M2, M2.stop (M2 ORF containing a translational stop codon at amino acid 13), or M2-mCherry upstream of the IRES-Thy1.1 cassette. (B) Primary B cells were isolated from naïve mice by negative selection and stimulated with LPS prior to retroviral transduction. Cell cultures were harvested at the indicated time points and cell surface Thy1.1 expression was analyzed by flow cytometry. (C) Supernatants from retrovirally transduced B cell cultures were analyzed for IL10 secretion by ELISA. Mean and standard deviation values for (B) and (C) were derived from compiling data from two independent experiments containing three replicates per condition per time point.