Fig 2: ox-LDL-induced foam cell formation and TRIM64 expression. THP-1 monocytes were primed with phorbol myristate acetate to induce macrophage differentiation. Subsequently, the macrophages were treated with or without (control) 50 mg/L ox-LDL for 48 h and then stained with A Oil Red O, and the B TC, C FC, D CE/TC levels in THP-1-derived macrophages were measured. Scale bars: 100 µm. E, F THP-1-derived macrophages were treated with 50 mg/L ox-LDL for 0, 12, 24, and 48 h, and then, TRIM64 expression was measured by quantitative RT-PCR and Western blot assays. *P < 0.05, **P < 0.01, ***P < 0.001 compared with control or 0 h. Data are expressed as mean ± SD of three biological replicates

Fig 3: TRIM64 overexpression promoted ox-LDL-induced foam cell formation, pyroptosis, and inflammation via the NF-?B signaling. THP-1-derived macrophages were transduced with TRIM64 expressing vectors (TRIM64) or blank vector (vector) and treated with 50 mg/L ox-LDL in the presence of 10 µM PDTC or vehicle for 48 h. A Cells were stained with Oil Red O. Scale bars: 100 µm. B TC, C FC, and D CE/TC levels were measured. E Western blot assays were performed to detect TRIM64, CD36, and ABCA1 expression. F Pyroptotic cell death was determined by flow cytometry. G, H Western blot assays were performed to detect NLRP3, ASC, caspase-1, and GSDMD-N expression. I IL-18, IL-1ß, TNF-a, and IL-6 content were measured by ELISA. J Western blot assays were performed to detect NF-?B expression in nucleus and cytoplasm. *P < 0.05, ***P < 0.001 compared with vector or vehicle + vector. #P < 0.05, ##P < 0.01, ###P < 0.001 compared with vehicle + TRIM64. Data are expressed as mean ± SD of three biological replicates

Fig 4: TRIM64 knockdown inhibited ox-LDL-induced foam cell formation, pyroptosis, and inflammation. THP-1-derived macrophages were transduced with TRIM64 silencing vectors (shTRIM64) or negative control (shNC) and treated with 50 mg/L ox-LDL for 48 h. A Cells were stained with Oil Red O. Scale bars: 100 µm. B TC, C FC, and D CE/TC levels were measured. E Western blot assays were performed to detect CD36 and ABCA1 expression. F, G Pyroptotic cell death was determined by flow cytometry. H Western blot assays were performed to detect NLRP3, ASC, caspase-1, and GSDMD-N expression. I IL-18, IL-1ß, TNF-a, and IL-6 content were measured by ELISA. J Western blot assays were performed to detect NF-?B expression in nucleus and cytoplasm. ***P < 0.001 compared with shNC. Data are expressed as mean ± SD of three biological replicates