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. Author manuscript; available in PMC: 2011 Nov 16.
Published in final edited form as: Cancer Cell. 2010 Nov 16;18(5):411–422. doi: 10.1016/j.ccr.2010.10.024

Figure 4. SAH-p53-8 blocks the formation of p53-HDMX complexes.

Figure 4

(A) JEG-3 choriocarcinoma cells were exposed to vehicle, 20 µM SAH-p53-8, or 20 µM Nutlin-3 for 6 h. Cellular extracts and anti-HDMX immunoprecipitates were subjected to SDS-PAGE and analyzed by western blotting for p53, HDMX, and HAUSP. (B) JEG-3 cells were treated with 0–20 µM SAH-p53-8 in the presence of 10 µM MG-132 for 6 h. Cellular extracts and anti-HDMX immunoprecipitates were analyzed by western blotting for p53, HDMX, and HAUSP. (C) Doxycycline-inducible U2OS cells were treated with or without doxycycline in the presence of Nutlin-3 (10 µM), SAH-p53-8 (10 µM), or both, and induction of HA-HDMX and p53 was detected and quantitated by immunofluorescence. Scale bar, 10 µm (D) Cultured U2OS cells were treated with or without doxycycline in the presence of Nutlin-3, SAH-p53-8, or both and then processed for P-LISA. Scale bar, 10 µm (E) Quantitation of p53-HDMX complexes as detected by P-LISA. Data are mean +/− s.d. *p=0.0003, **p=0.0001, unpaired t-test with Welch’s correction.