Figure 7.

Injection of lenti-TGFB3-AS1 accelerates inflammation induced by Hcy in CBS^+/− mice

(A) qRT-PCR analyzed the TGFB3-AS1 expression in peripheral blood monocytes of CBS^+/− mice fed with the high-methionine diet after 4-week injection of lenti-TGFB3-AS1, lenti-GFP, or PBS. (B) The contents of proinflammatory cytokines IL-6, IL-1β, and TNF-α in serum of CBS^+/− mice injected with lenti-TGFB3-AS1, lenti-GFP, or PBS were measured using an automatic biochemical analyzer. (C) The monocyte populations in BM, peripheral blood, and spleen isolated from CBS^+/− mice after injection of lenti-TGFB3-AS1, lenti-GFP, or PBS were analyzed by flow cytometry. CD11b⁺ and Ly-6C⁺ monocyte subsets were further characterized using anti-CD11b and anti-Ly-6C antibodies. SSC, side scattered light; FSC, forward-scattered light. (D) The percentages of mononuclear cell (MNC) and CD11b⁺Ly-6C⁺ inflammatory monocyte subsets in MNC were analyzed. (E) The expression levels of miR-144 in peripheral monocytes from CBS^+/− mice injected with lenti-TGFB3-AS1, lenti-GFP, or PBS were detected by qRT-PCR. (F–H) The expression levels of Rap1a and Wnt signaling pathway-related proteins (wnt3a, wnt5a, β-catenin) in peripheral monocytes from CBS^+/− mice injected with lenti-TGFB3-AS1, lenti-GFP, or PBS were analyzed by qRT-PCR and western blot. ∗p < 0.05, ∗∗p < 0.01, compared with lenti-GFP group.