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. Author manuscript; available in PMC: 2014 Jan 15.
Published in final edited form as: J Immunol. 2012 Dec 14;190(2):756–763. doi: 10.4049/jimmunol.1201811

Figure 2. MMSET facilitates CSR. A.

Figure 2

CH12F3 cells were infected with lentivirus expressing the indicated shRNA, and CSR was assessed using flow cytometry analysis of IgM and IgA expression following CIT. Numbers indicate the percentage of total live IgA+ B cells. The right panels show the knockdown efficiency of 53BP1and MMSET by western blot. Data are representative of two independent experiments. B, Evaluation of the effect of MMSET expression on cell growth. Cell numbers of CH12F3 cells expressing either control shRNA or different MMSET-specific shRNAs were counted on 3 consecutive days. C, Evaluation of the effect of MMSET expression on cellular viability of pre-switched CH12F3 cells. Similar to the experiments in B, cell viability of CH12F3 cells expressing either control shRNA or different MMSET-specific shRNAs were analyzed on 3 consecutive days. D, Evaluation of the effect of MMSET expression on cell viability of post-switched CH12F3 cells. Class switched IgA+/IgM-CH12F3 cells expressing either control or MMSET-specific shRNA were sorted, and cell viability was analyzed. E, Western analysis of AID expression in CH12F3 cells pre- and post-MMSET knock down by specific shRNAs. F, ChIP analysis of γH2AX at the Sμ1 region and Sα region of Igh locus in CH12F3 cells transfected with the indicated shRNAs, with or without CIT stimulation. Three independent experimental replicates were performed for each experiment (± s.e.m., n=3).