Fig. 6: The ribosome stress is triggered by NAF1 depletion in glioma cells. | Oncogenesis

Fig. 6: The ribosome stress is triggered by NAF1 depletion in glioma cells.

From: Increased expression of NAF1 contributes to malignant phenotypes of glioma cells through promoting protein synthesis and associates with poor patient survival

Fig. 6

a Western blot analysis of MDM2 and p53 proteins upon NAF1 knockdown in U87 and SHG44 cells. Using GAPDH and Actin as loading controls, and the western blot is representative of three independently preformed experiments. b, c Western blot analysis was conducted to evaluate the effect of the NAF1 depletion on the expression and cellular localization of the NPM1 and RPS14 in U87 and SHG44 cells. Using GAPDH, Actin and histone H3 as loading controls, and the western blot is representative of three independently preformed experiments. d The effect of NAF1 knockdown on subcellular localization of RPS14 and NPM1 in U87 cells was investigated by the immunofluorescence assay. Red color represents nuclear and cytoplasmic staining of RPS14; green color represents nuclear and cytoplasmic staining of NPM1; blue color represents Hoeschst 33342 staining for nuclei; fluorophores in the three-color overlay are labeled to the rightmost. Actinomycin D treatment was used as positive control, while DMSO treatment was used as the vehicle control. Scale bars, 500 μm. The experiments were performed in triplicate. e Working model of NAF1 depletion triggering ribosome stress. Briefly, upon NAF1 reduction, NPM1 is dissociated from nucleolar to nucleoplasm. As well, the waved circle indicates the RPS14 which is released from breaking down of cytoplasmic or nucleolar ribosomes, entering the nucleoplasm to interact with MDM2. This will result in p53 reactivation by blocking MDM2-p53 feedback loop

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