Figure 4 | Cell Death & Disease

Figure 4

From: Raf kinases mediate the phosphorylation of eukaryotic translation elongation factor 1A and regulate its stability in eukaryotic cells

Figure 4

Ubiquitination assay of eEF1A1-HIS and eEF1A2-HIS and their mutants. After cotransfection of COS 7 cells with wt eEF1A-HISs and corresponding mutants with HA-ubiquitin wt and treatment of cells in the absence (−) or in presence (+) of MG132, cell extracts were analyzed after pull-down for eEF1A1-HIS (a, b, i) and eEF1A2-HIS (e, f, k), respectively, by immunoblotting with anti-HA (a, b, e, f) and anti-His (i, k) antibodies. (a, b, i) Lanes: 1, eEF1A1-HIS wt; 2, eEF1A1-HIS S21A; 3, eEF1A1-HIS S21D; 4, eEF1A1-HIS T88A and 5, eEF1A1-HIS T88D. (e, f, k) Lanes: 1, eEF1A2-HIS wt; 2, eEF1A2-HIS S21A and 3, eEF1A2-HIS S21D. Band intensities were analyzed using Image J software (National Institutes of Health, Bethesda, MD, USA) (c, d, g, h, j and l)

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