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. 2021 Feb 3;11:2967. doi: 10.1038/s41598-021-82542-4

Figure 5.

Figure 5

Prolonged MDM4 inhibition with ATSP-7041 and NSC207895 lead to upregulation of p53 activity and downstream signaling. (a) Bar graphs representing normalized mRNA expression of p53 targets Bax, Puma, CDKN1A, and MDM2 analyzed with qPCR experiments. RNA extracted from HepG2 and HepT1 cells treated with the indicated concentrations of ATSP-7041 for 48 h was compared to that from untreated cells (0 h). Error bars represent SD. Data shown are representative of at least three independent experiments performed with three replicate wells each time. Student’s t test (two tailed) *P < 0.05, **P < 0.01, ***P < 0.001. (b) Bar graphs representing normalized mRNA expression of p53 targets Bax, Puma, CDKN1A, and MDM2 analyzed with qPCR experiments. RNA extracted from HepG2 and HepT1 cells treated with the indicated concentrations of NSC207895 for 48 h was compared to that from untreated cells (0 h). Error bars represent SD. Data shown are representative of at least three independent experiments performed with three replicate wells each time. Student’s t test (two tailed) *P < 0.05, **P < 0.01, ***P < 0.001. (c) Protein lysis from cells treated with 0.5 μM ATSP-7041 or NSC207895 for 48 h were compared to untreated, control cells (0 h). Immunoblotting was done with the indicated antibodies, including MDM4 (04–1555, Millipore). β-Actin immunoblotting was used as a loading control. Data shown are representative of at least three independent experiments. Full length blots are presented in Supplementary Fig. 9.