Fig. 6: FMDV VP4 induces NME1 reduction to promote interaction of p53 with MDM2. | Cell Death & Disease

Fig. 6: FMDV VP4 induces NME1 reduction to promote interaction of p53 with MDM2.

From: Foot-and-mouth disease virus induces lysosomal degradation of NME1 to impair p53-regulated interferon-inducible antiviral genes expression

Fig. 6

a HEK-293T cells grown in 10-cm dishes were co-transfected with 6 μg Flag-MDM2 plasmid with the indicated plasmids (6 μg), and MG132 was added at 20 μM. The cells were collected and lysed at 24 hpt and immunoprecipitated with HA antibody. Expression of Flag-MDM2 and HA-p53 was detected using appropriate antibodies. Relative fold-change in abundance of Flag-MDM2 was determined by densitometric analysis. Whole cell lysates were also analyzed by Western blotting using appropriate antibodies. b HEK-293T cells grown in 10-cm dishes were co-transfected 6 μg Flag-MDM2 plasmid, 6 μg HA-p53 plasmid, 6 μg Myc-vector or Myc-NME1 plasmids, and 6 μg Flag-vector or different doses of Flag-VP4 plasmids (0, 3, or 6 μg). MG132 was added at 20 μM. The cells were collected and lysed at 24 hpt and immunoprecipitated with HA antibody. Expression of Flag-MDM2 and HA-p53 was detected using appropriate antibodies. Relative fold-change in abundance of Flag-MDM2 was determined by densitometric analysis. Whole cell lysates were also analyzed by Western blotting using appropriate antibodies. All the experiments were repeated three times

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