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Figure 3


Fig. 3. POT1A interacts with TERT-V(I8) in vitro. The presence (+) or absence (–) of particular components is indicated above: myc, translation mix using control vector, pGBKT7 as template; myc-POT1A, translation mix using POT1A cloned in pDEST-GBKT7; and [35S-met]-HA-TERT-V(I8), translation mix using TERT-V(I8) cloned in pDEST-GADT7. The upper panel [labelled AtTERT V(I8)] is an autoradiograph revealing the presence or absence of the TERT fusion protein. The lower panel (labelled POT1A) is a western blot revealing the presence or absence of the POT1A fusion protein. Heavy chain IgGs are indicated by arrowheads. Lane 1, radiolabelled [35S-met]-TERT-V(I8) synthesis reaction; lane 2, POT1A synthesis reaction (myc-tagged); lanes 3 and 4, input: the mixed proteins after an overnight incubation with antibodies and protein A beads, correspond to 0.2% of the total amount of protein used for immunoprecipitation; lanes 5 and 6, IP anti-myc: the proteins remaining bound to the protein A beads after immunoprecipitation.





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