Panel A: Western blot analysis validating the TET2 antibody in transduced and non-transduced HEK293T cells. The upper section shows flow cytometry plots for control and TET2-Flag–transduced cells, while the lower section presents western blot results with lanes corresponding to different cell conditions alongside a protein ladder. Panel B: TET2 immunoprecipitation performed on day 3 and day 6 samples, followed by immunoblot detection of TET2 and AID (Activation-Induced Cytidine Deaminase). Panel C: Repeat TET2 immunoprecipitation on day 3 and day 6 samples, with immunoblot analysis for AID. Panel D: AID immunoprecipitation on day 3 and day 6 using an anti-AID antibody, followed by immunoblot detection of AID and TET2 (Ten-Eleven Translocation methylcytosine dioxygenase 2). Panel E: AID immunoprecipitation on day 3 and day 6 using an alternative anti-AID antibody, with immunoblot detection of AID and TET2. Panel F: DNMT1 (DNA methyltransferase 1) immunoprecipitation on day 3 and day 6 samples, followed by immunoblot detection of AID and DNMT1.
Co-immunoprecipitation of AID, TET2, and DNMT1 in iGB cells. (A) Validation of TET2 antibody by western blot in TET2-transduced and nontransduced cells. Upper: FACS plot for the control and TET2-Flag–transduced HEK293T cells. Lower: Western blot to detect TET2. Lane 1: Nontransduced HEK293T cells; lanes 3–6: WT and AIDR112H iGB cells underwent TET2 transfection by AMAXA electroporation; Lane 7: protein ladder. (B) TET2 immunoprecipitation on day 3 and day 6. Immunoblot for TET2 (Abcam, upper panel) and AID (Active Motif, lower panel), cropped blot images shown in Fig. 8 E. (C) Repeat of TET2 immunoprecipitation on day 3 and day 6. Immunoblot for AID. (D) AID immunoprecipitation on day 3 and day 6 using anti-AID antibody (Active Motif). Immunoblot for AID (Active Motif, upper panel) and TET2 (Abcam, lower panel). (E) AID immunoprecipitation on day 3 and day 6 using anti-AID antibody (eBioscience). Immunoblot for AID (Active Motif, left panel) and TET2 (Abcam, right panel). (F) DNMT1 immunoprecipitation on day 3 and day 6. Immunoblot for AID (Active Motif) and DNMT1 (Merck). Source data are available for this figure: SourceData FS4.