Figure 4.

FBXL15 possess a destabilizing C-terminus. N- or C-terminus-HiBiT-tagged FBXL15 turnover was measured by CHX assay with transient expression of FBXL15 fusion variants and co-treatment with 50 μg/ml CHX for 4 h and, where indicated, 20 μM MG132 proteasome inhibitor. Protein levels were measured and normalized to the number of living cells as described in Materials and Methods; assays were normalized to the corresponding measurement for each protein variant from control time 0′. Error bars denote the standard deviation from the mean derived from three biological replicates; dots represent the biological replicates. Each biological replicate is a mean of three technical replicates. Data was analyzed using two-way ANOVA and the significance levels obtained from Tukey's multiple comparisons test are indicated for the compared conditions (∗∗∗∗ P ≤ 0.0001; ∗∗ P ≤ 0.01).