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. 2024 Dec 17;25(24):13524. doi: 10.3390/ijms252413524

Figure 6.

Figure 6

ITCH mediates FTL degradation in endothelial cells. (A) mRNA expression of FTL in HAECs that were incubated with 100 μg/mL ox-LDL at various times. (B) HAECs pretreated with 3-methyladenine (3-MA, 5 mM), chloroquine (25 μM), or MG132 (20 μM) for 1 h were incubated in the presence and absence of ox-LDL (100 μg/mL) for 24 h. Then, protein expression of FTL was determined. (C) HAECs transfected with si-ITCH or si-con in the presence of cycloheximide (CHX, 100 μg/mL) for up to 8 h. Protein expression of FTL was determined. For (A–C), n = 6 independent experiments. (D) The ubiquitination of FTL in ITCH siRNA-transfected HAECs under ox-LDL (100 µg/mL) for 24 h in the presence of MG132 (20 µM). (E,F) HAECs were incubated with 100 μg/mL ox-LDL in the presence of MG132 (20 μM) for 24 h, and cell lysates were then immunoprecipitated to verify endogenous interaction between ITCH and FTL. Results are presented as the mean ± S.D. # p < 0.05 vs. control group, * p < 0.05 vs. ox-LDL alone group.