Supplementary Figure 7: Binding of TERB1 to TRF2 in vitro.
From: Dissecting the telomere–inner nuclear membrane interface formed in meiosis
(a) Direct binding curves of biotin-tagged TRF1TRFH and TRF2TRFH with Alexa Fluor 488-labeled GST-TERB1TBM. Mean of technical duplicate is plotted. Mean and s.e.m. of duplicate measurements are indicated. (b) Competition experiments demonstrating similar affinity of TRF1 and TRF2 for TERB1TBM peptide. Mean of technical duplicate is plotted. Error bars indicate s.e.m. (c-d) Fluorescence-based competition experiments using Alexa Fluor 488-labeled GST-TERB1TBM pre-bound to biotin-tagged TRF2TRFH on streptavidin beads titrated with varying concentrations of indicated unlabeled TERB1TBM peptides (c) or TERB1TRFB – TERB21-107 complexes (d). For c and d, mean of technical duplicate is plotted and two biological replicates were performed. (e) Ternary pull down on amylose resin showing that TRF2 associates with MBP-tagged TERB1TRFB – TERB21-107 in the absence of Rap1 but not in its presence. (f) Ternary pull down on amylose resin showing that TRF2 associates with MBP-tagged TERB1TRFB – TERB21-107 in the presence or absence of TIN2.
