Journal: Journal of Biomedical Science

Article Title: Smooth muscle-specific expression of hydroxyindole O-methyltransferase reduces arterial injury-induced intimal hyperplasia

doi: 10.1186/s12929-025-01172-4

Figure Lengend Snippet: Serotonin reduces VSMC proliferation and marker expression via the ERK1/2 pathway. A Serum-starved wild-type (WT) VSMCs were plated, wounded, and then treated with vehicle, different doses of serotonin, or IL-1β (10 ng/mL, as positive control) in starvation medium for 6 h. Wound images were captured at time 0 and at 6 h. Wound closure was quantified, and % wound closure was calculated (n = 4 each). B WT VSMCs were stimulated with different doses of serotonin for 24 h in the presence or absence of IL-1β (2 ng/mL), and proliferation was assessed by CCK-8 assays. n = 3 each. C Serum-starved WT and transgenic (TG) VSMCs were stimulated with vehicle, IL-1β (2 ng/mL), or 25 µmol/L serotonin + IL-1β (2 ng/mL) for 24 h, and proliferation was assessed. n = 4 each. D WT VSMCs were treated with vehicle, serotonin, or serotonin in the presence of 5-MTP (100 µmol/L). Proteins were prepared 24 h later for Western blotting to detect SM-MHC, SM22α, SM α-actin, and α-tubulin as a loading control. Quantitative analysis of VSMC markers, n = 4 each group. E WT and TG VSMCs were stimulated with vehicle, serotonin, or IL-1β (10 ng/mL) for 15 min, and proteins were prepared for Western blotting to detect phosphorylated and total ERK1/2 and p38, hHIOMT, and α-tubulin as a loading control. Quantitative analysis of ERK1/2 activation in WT and TG VSMCs. n = 3 for each group. F WT VSMCs were treated with vehicle or serotonin in the absence or presence of U0126. Proteins were prepared 24 h later for Western blotting to detect VSMC markers and α-tubulin as a loading control. Quantitative analysis of VSMC markers, n = 4 for each group. Different letters indicate significant differences between groups as determined by one-way ANOVA followed by Tukey's test

Article Snippet: To detect SMC markers, blots were incubated with primary antibodies against SM 22α (smooth muscle 22 alpha) (Abcam; ab14106), SM α-actin (smooth muscle alpha-actin) (Sigma-Aldrich; A5228), and SM-MHC (smooth muscle myosin heavy chain) (Proteintech, Rosemont, IL, USA; 21404–1-AP).

Techniques: Marker, Expressing, Positive Control, CCK-8 Assay, Transgenic Assay, Western Blot, Control, Activation Assay

Journal: Experimental Biology and Medicine

Article Title: Mechanisms of AGE-induced VSMC phenotypic switching and macrophage modulation in human abdominal aortic aneurysms

doi: 10.3389/ebm.2025.10527

Figure Lengend Snippet: AGE Effects on Human VSMCs: Promotion of Migration and Inhibition of Contraction. (A) ELISA analysis detected the accumulation of AGEs in lysates of human aortic tissues. (B–D) RT-qPCR (B,C) and Western blot (D) detected the expression of the contraction marker α-SMA and the migration marker MMP-2 in human VSMCs after AGE treatment for 24 h; (E) Transwell migration experiment detected the migration of human VSMCs after AGE treatment for 24 h or 72 h; (F) Immunofluorescence detected the expression of the contraction marker MYH11 in human VSMCs after AGE treatment for 24 h. Blue fluorescence indicates DAPI staining, and green indicates MYH11 staining. (G) The CTG assay detected the growth curve of human VSMCs under the treatment with 200 μg/mL AGE. Data are presented as the mean ± SD from three independent experiments and were analyzed with a one-way ANOVA followed by a Dunnett’s multiple comparisons test. Asterisks indicate significant differences (*P < 0.05, **P < 0.01, ***P < 0.001) between the treatment group and the control group.

Article Snippet: After blocking with 3% BSA at room temperature for 30 min, the cells were incubated overnight at 4°C with MYH11 Antibody (1:100, Proteintech, Cat. No. 21404-1-AP).

Techniques: Migration, Inhibition, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Expressing, Marker, Immunofluorescence, Fluorescence, Staining, CTG Assay, Control