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. 2012 Jun 26;21(19):4225–4236. doi: 10.1093/hmg/dds246

Figure 3.

Figure 3.

Interactions between PTIP and poly-Q AR. (A) PC12 cells stably expressing AR10Q or AR112Q were induced with synthetic ligand R1881 and protein lysates immunoprecipitated with anti-AR antibodies as indicated. Addition of ligand induced a slower migrating PTIP isoform (input) in AR112Q cells. The AR112Q protein binds to this slower migrating form of PTIP, whereas AR10Q shows no interaction. (B) The slower migrating form of PTIP is specific for AR112Q PC12 cells after the addition of ligand (arrows). The presence of the slower PTIP isoform is inhibited by the DNA-PKcs inhibitor Nu7026 but not by the ATM inhibitor Ku55933. (C) Lambda phosphatase reduces the slower migrating form of PTIP in AR112Q-expressing cell lysates. (D) Western blot of lysates from transient expression of AR24Q and AR113Q in HEK293 cells. Note slower migrating PTIP isoform is not radiation inducible and only appears when poly-Q AR is expressed. (E) AR112Q induces ATM phosphorylation in PC12 cells and higher levels of γ-H2AX even in the absence of IR. 5 Gy IR activates P-ATM and increases γ-H2AX in both cell types. (F) Western blot of lysates from AR10Q and AR112Q PC12 cells showing p53 phosphorylation in response to IR. Note, P-p53 levels in AR112Q cells are elevated prior to radiation.