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. 2021 Aug 30;29(2):323–336. doi: 10.1038/s41418-021-00855-3

Fig. 3. Bmf T72 phosphorylation disrupts Bmf/DLC2 binding.

Fig. 3

A Top: schematic show of DLC2 binding domain of Bmf protein; Bottom: SDS PAGE of purified 6xHis-DLC2 protein. B–F ITC analysis of interaction between 6xHis-DLC2 and non-phosphorylated (B), pS74 (C), pS77 (D), pS74/pS77 (E) or pT72 (F) Bmf peptides. G 293 T or MCF-7 cells were co-transfected with HA-Bmf∆C26 or FLAG-DLC2 expressing plasmids for 48 h. Cells were lysed, and reciprocal co-immunoprecipitations were performed using HA- or FLAG-tag antibodies with IgG being the negative control. H Lentivirus-transduced MCF-7 or MCF-10A cells inducibly expressing full length HA-Bmf variants (WT, T72E, S74E and S77E) were induced with doxycycline (100 ng/mL) for 4 h and lysed for immunoprecipitation using HA antibody. Immunoprecipitants were analyzed by HA and DLC2 antibodies.