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. 2024 Jan 11;134(5):e166149. doi: 10.1172/JCI166149

Figure 5. TRIM56 interacts with FASN.

Figure 5

(A and B) Interaction of FLAG-TRIM56 with HA-FASN in HEK293T cells as demonstrated by IP (n = 3). (C) HA-FASN interacted with endogenous TRIM56 in HEK293T (n = 3). (D and E) GST-pulldown assay confirmed the interaction between TRIM56 and FASN in HEK293T cells (n = 3). (F) FASN-TRIM56 direct interaction as determined by SPR analysis. (G) Molecular characterization of FASN interaction with different truncated fragments of TRIM56 in HEK293T cells (n = 3). (H) Determination of intracellular TG content in HepG2 cells transfected with empty vector (FLAG) or FLAG -TRIM56 or the FLAG-TRIM56 (1–521 aa) fragment in the presence of PO (n = 6). ***P < 0.001, by 1-way ANOVA followed by Bonferroni’s post hoc test. (I) HepG2 were transfected with empty vector or FLAG-TRIM56, or FLAG-TRIM56 (1–521 aa) fragment in the presence or absence of PO before Nile Red staining (n = 3). Scale bars: 50 μm. (J) HepG2 cells were transfected with empty vector (FLAG) or FLAG-TRIM56 (1–521 aa) in the presence of PO before whole-cell lysate was collected for Western blotting to determine the expression of proteins related to fatty acid metabolism (n = 3). (K) Expression of the indicated genes in HepG2 cells transfected with empty vector (FLAG) or FLAG-TRIM56 (1–521 aa) in the presence of PO (n = 5). NS, by 2-tailed Student’s t test for SCD1 and DGAT2 and Mann-Whitney U test for ELOVL1, ELOVL6, and GPAM.