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

Figure 6.

Figure 6

14-3-3 antagonist Difopein decreases the stability of WT UL97.A, U-2 OS cells were transfected with an expression vector for WT or S13A mutant UL97 (0.4 μg) and the indicated Flag-tagged 14-3-3 isoform (0.6 μg) together with the indicated amounts (μg) of an expression plasmid for EYFP fused to Difopein. Plasmid amounts in transfections were equalized with EYFP only expression plasmids. After 48 h, cells were harvested and lysates were subjected to Western blotting with the indicated antibodies. B, the transfection with either no (white bar) or 0.3 μg (gray bar) of an expression plasmid for EYFP fused to Difopein shown in panel (A) were performed in biological triplicates. The protein level of HA-tagged UL97 was quantified and normalized to GAPDH. Values are presented relative to the value in the absence of EYFP fused to Difopein for 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; ∗∗p < 0.01; n.s., not significant (p > 0.05).