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. Author manuscript; available in PMC: 2017 Oct 10.
Published in final edited form as: Cancer Cell. 2016 Sep 22;30(4):623–636. doi: 10.1016/j.ccell.2016.08.015

Figure 5.

Figure 5

Effect of leads on expression and function of p53 and XIAP following inhibition of MDM2 (A) Western blot for expression of p53 and its targets p21 and PUMA in EU-1 cells treated with 1 μM MX3 or 10 μM MX69 for indicated times. (B) Cell-cycle distribution of EU-1 and EU-1/sip53 cells treated with MX69 for 8 hr. (C) Representative flow cytometry histographs for EU-1 cells, both control and treated with 1 μM MX3 and 10 μM MX69. (D) Western blot for expression of MDM2 and XIAP in SH-EP1 cells with MDM2 KO by CRISPR/Cas9 (sgMDM2), treated with indicated doses of MX69 for 24 hr. (E) EU-1 cells were treated with either 10 μM MX69 or MX68 (control) for 4 hr and their cytoplasmic lysates were fractionated on a sucrose gradient. RNA was extracted from each of the fractions and subjected to qRT-PCR for analysis of the distribution of XIAP and GAPDH mRNAs. Data show the percentage of the total amount of corresponding mRNA in each fraction and represent mean ± SD of three independent experiments. (F) ELISA for activation of caspase-7 and -9 in ALL and NB cells treated with different doses of MX69 as indicated for 8 hr. (G) Western blot showing activation of caspase-3 and cleavage of death substrate PARP in cancer cells following treatment with 10 μM MX69 and MX68 (control) or 1 μM MX3 for times indicated. Data in (B and F) represent mean of three independent experiments, bars ± SD. See also Figure S5.