Fig. 4: ATO causes ribosome stalling and p97 inhibitor increases the sensitivity of APL to ATO. | Cancer Gene Therapy

Fig. 4: ATO causes ribosome stalling and p97 inhibitor increases the sensitivity of APL to ATO.

From: Arsenic trioxide and p97 inhibitor synergize against acute myeloid leukemia by targeting nascent polypeptides and activating the ZAKα–JNK pathway

Fig. 4

A Polysome profiles from NB4 cells with or without 1.5 μM ATO treatment for 6 h. B Immunoblot of indicated proteins in the NB4 cells at 0, 4, 20 h with 0.5 or 2 μM ATO treatment. C NB4 cells were pre-treated with 5 μM JNK inhibitor (JNK-IN-8) for 1 h, and then 2.5 μM ATO was added for another 24 h. Apoptosis was detected by flow cytometry. D Immunoblot of ZAKα and p-ZAKα in the NB4 cells at 4 or 20 h with 0.5 or 2 μM ATO treatment. The phosphorylated proteins in the cell lysate were enriched by Phos-tagTM Agarose. E The apoptosis ratio of ZAKα-knockdown NB4 cells with or without ATO treatment for 24 h. F Immunoblot of ZAKα in the ZAKα-knockdown NB4 cells. G The NB4 cells were treated with 1 μM Biotin-As for 4 h, and then cell fractions were prepared for the Co-IP assays using the streptavidin magnetic beads and indicated antibodies. H The NB4 cells were treated with 1 μM Biotin-As for 4 h, and then 20 μM puromycin was added for another 30 min. Cell fractions were prepared for the Co-IP assays using the NEMF, p97, and puromycin antibodies. I The NB4 cells were pre-treated with or without 1 μM NMS873 for 1 h and then indicated doses of ATO were added for another 24 h. The apoptosis ratio was detected by flow cytometry. Two-tailed Student t test, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Error bars reflect ± SEM of three independent experiments.

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