(A) Representative immunoblots of endogenous nuclear PPARγ
interactions with MRN and UBR5 at baseline and upon DNA damage induced by HU and
doxorubicin (DoxR).
(B) Representative immunoblots of HU-induced pATM and its targets with
PPARγ or UBR5 depletion.
(C) Representative immunoblots of reduced UBR5 binding to ubiquitinated
proteins with PPARγ depletion.
(D and E) Representative immunoblots of ATMIN and pRPA2 levels with
PPARγ (D) or UBR5 (E) depletion upon HU treatments.
(F) Cells were transfected with HA-tagged ubiquitin and subsequently the
siRNA as indicated. Cells were treated with the proteasome inhibitor MG132 (MG)
for 2 h before lysis in a denaturing buffer. Endogenous ATMIN was
immunoprecipitated to determine its polyubiquitinated form. Representative
immunoblots show effects of PPARγ or UBR5 depletion on endogenous ATMIN
ubiquitination detected by anti-hemagglutinin (HA) antibody.
(G) Quantitative real-time PCR shows effects of PPARγ or UBR5
depletion by the respective siRNA on ATMIN mRNA levels
(normalized to β-actin mRNA).
siC, siControl; siPg, siPPARγ; siU5, siUBR5; Veh; vehicle. Error
bars, mean ± SEM.
See also Figure
S3.