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. 2022 Sep 20;298(11):102513. doi: 10.1016/j.jbc.2022.102513

Figure 5.

Figure 5

Serine 13 contributes to the protein stability of transfected UL97.A, U-2 OS cells were transfected with an expression vector for WT UL97 (0.6 μg) or the S13A mutant of UL97 (1.2 μg) together with expression vectors for the indicated Flag-tagged 14-3-3 isoform. Plasmid amounts in transfections were equalized with empty vectors. After 48 h, cells were treated with 50 μg/ml of cycloheximide (CHX) for the indicated times, and harvested lysates were subjected to Western blotting with the indicated antibodies. A representative image from one of between three and six independent biological replicates is shown. B, the protein level of HA-tagged UL97 from the experiments shown in panel (A) was quantified and normalized to GAPDH protein. Values are presented relative to the value in 0 h after CHX treatment of each transfection of WT or S13A-UL97 (set at 100%). Error bars denote SDs. Statistical analysis utilized a two-tailed unpaired Student’s t test. ∗p < 0.05; n.s., not significant (p > 0.05).