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. 2025 Jan 7;8:0574. doi: 10.34133/research.0574

Fig. 2.

Fig. 2.

tRF-AspGTC overexpression induces VSMC phenotypic switching and inflammation. (A) RT-qPCR analysis of overexpression efficiency following transfection with a tRF-AspGTC mimic; n = 3 per group. (B) Western blot (WB) analysis of contractile VSMC marker proteins myosin heavy chain (MHC), calponin 1 (CNN1), and α-SMA expression levels after tRF-AspGTC overexpression. (C) Quantitative analysis of (B); n = 3 per group. (D) WB analysis of matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP9) protein expression levels. (E) Quantitative analysis of (D); n = 3 per group. (F) WB analysis of MHC, CNN1, and α-SMA expression levels in VSMCs treated with 400 μM hydrogen peroxide for 24 h after tRF-AspGTC overexpression. (G) Quantitative analysis of (F); n = 3 per group. (H) WB analysis of MMP2 and MMP9 protein expression levels. (I) Quantitative analysis of (H); n = 3 per group. (J) RT-qPCR analysis of MMP2 and MMP9 messenger RNA (mRNA) levels following tRF-AspGTC overexpression; n = 3 per group. (K) RT-qPCR analysis of inflammatory cytokines interleukin 1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), and interleukin 6 (IL-6) mRNA expression levels; n = 3 per group. (L) Measurement of reactive oxygen species (ROS) generation in VSMCs treated with hydrogen peroxide following tRF-AspGTC overexpression. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. NC, negative control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.