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. 2017 Jul 19;45(16):9455–9466. doi: 10.1093/nar/gkx621

Figure 4.

Figure 4.

POLA2-binding by STN1 variants. (A) A schematic depiction of the tagged STN1 and POLA2 proteins used for the pull down assays is shown. (B) Purified, Strep-tagged STN1F, STN1N, STN1C and STN1N mutants were bound to Strep-Tactin beads. Purified POLA21–414-FG3 was used as the prey in the Strep-Tactin pull down/elution experiments. The elutions were analyzed by western blot to assess the binding of POLA2 to STN1 variants and mutants. The level of POLA2 was measured by probing the blot with anti-FG antibody, while that of STN1 by anti-Strep antibody. (C) The ratio of POLA2 to STN1 in the elution samples was calculated, normalized against the value for wild-type STN1N and plotted. Data (averages ± S.E.M) are from three or more independent experiments. P-values as determined by two-tailed t-test are designated by asterisks as follows: **: 0.1 > P > 0.05; ***: 0.05 > P > 0.01; ****: 0.01 > P.