Extended Data Fig. 2: FAM72A interacts with UNG2 and Fam72a-/- CH12 cells have no defects in proliferation, AID expression or switch region transcription.
From: Fam72a enforces error-prone DNA repair during antibody diversification
a, Western blot analysis for Flag, UNG (UNG1 and UNG2) and β-Actin in wildtype and Fam72a-/- CH12 cells transduced with a retrovirus expressing FAM72A-Flag or FAM72AW125A-Flag before (Input) or after immunoprecipitation with a Flag-specific antibody (Pulldown). b, Schematic representation of the murine Fam72a locus and location of the gRNAs used to generate Fam72a-/- clones using CRISPR/Cas9-HF1. c, RT-PCR analysis for Fam72a and Igβ in Fam72a+/+ and Fam72a-/- CH12 cells cultured or not for 3 days with TGFβ, anti-CD40 antibody and IL-4 (CIT). Data are representative of three experiments. d, Western blot analysis for AID and β-actin in wild-type (pCH12) and Fam72a-/- CH12 cells cultured or not for 3 days with TGFβ, anti-CD40 antibody and IL-4 (CIT). e, CFSE dye-dilution analysis by flow cytometry in Fam72a+/+ and two independent Fam72a-/- CH12 cell clones cultured for 3 days with TGFβ, IL-4 and anti-CD40 antibody. Data are representative of three experiments. f, RT-qPCR analysis for GLTμ and GLTα in Fam72a+/+ and Fam72a-/- CH12 cells cultured for 3 days with TGFβ, IL-4 and anti-CD40 antibody. Triplicates were normalized to the abundance of Igβ and are expressed relative to Fam72a+/+, set as 1. Statistical significance was determined by a two-tailed Student’s t-test. Data are presented as mean ± s.d. and are representative of 3 experiments. For gel source data, see Supplementary Fig. 1
