Figure 3. Regulation of RPA displacement by TERRA.

a, NEs (34 ng/μl) were treated with increasing concentrations (2, 4, 10, 20 nM) of TERRA (UUAGGG)₃, control RNA (CCCUAA)₃, or mock treated. The treated NEs were then incubated with RPA-coated ssTEL (2 nM), and the remaining RPA2 on ssTEL was analyzed after the incubation. b, The RPA displacing factors were captured with RPA-ssTEL as in Fig. 2a. The elution was incubated with TERRA or control RNA, and then applied to RPA-ssTEL. c, Purified hnRNPA1 (4.8 nM) was incubated with increasing concentrations of TERRA or control RNA (2, 4, 10, 20 nM), and then incubated with RPA-ssTEL (0.8 nM). d, hnRNPA1-coated ssTEL (0.8 nM) was incubated with increasing concentrations of TERRA (2, 20, 200, 2000 nM). The remaining hnRNPA1 on ssTEL was analyzed by Western blot. e, hnRNPA1-coated ssTEL (0.8 nM) was incubated with increasing concentrations of TERRA (2, 20, 200 nM) in the presence of POT1-TPP1 (2.4 nM). The hnRNPA1 and POT1 on ssTEL were analyzed by Western blot. f, A model for RPA displacement.