Journal: The Journal of Biological Chemistry

Article Title: Farnesyl pyrophosphate synthase promotes restenosis after vascular injury by activating small G proteins

doi: 10.1016/j.jbc.2025.110861

Figure Lengend Snippet: Lenti-FPPS shRNA-mediated knockdown of FPPS attenuates MAPK pathway activation induced by Ang II or TGF-β1 in VSMCs. A , representative Western blots showing phosphorylation levels of p38, ERK, and JNK in VSMCs under different treatments. Total p38, ERK, JNK, and β-actin were used as internal loading controls. B , quantitative analysis of the phosphorylation ratios (p-p38/p38, p-ERK/ERK, and p-JNK/JNK) normalized to β-actin and expressed as fold change relative to the normal control (NC) group. VSMCs were pretreated with Lenti-FPPS shRNA (MOI = 50, 48 h) followed by stimulation with Ang II (10 -7 mol/l) or TGF-β1 (10 ng/ml) for 24 h. Data are presented as mean ± SD from three independent experiments. Statistical analysis was performed by one-way ANOVA with Tukey’s post hoc test. ∗∗ p < 0.01 versus NC group; # p < 0.05 versus Ang II or TGF-β1 group. Ang II, angiotensin II; ERK, extracellular signal–regulated kinase; FPPS, farnesyl pyrophosphate synthase; JNK, Jun NH2-terminal kinase; MAPK, mitogen-activated protein kinase; MOI, multiplicity of infection; TGF-β1, transforming growth factor-beta 1; VSMC, vascular smooth muscle cell.

Article Snippet: The membranes were incubated overnight with the primary antibodies as follows: FPPS (Abcam; catalog no.: ab153805, 1:2000 dilution), CTGF (Abcam; catalog no.: ab6992, 1:1000 dilution), GFP (Abcam; catalog no.: ab183734, 1:1000 dilution), extracellular signal–regulated kinase (ERK; BOSTER Biological Technology Co, Ltd; catalog no.: BM4326, 1:1000 dilution), phospho-ERK (p-ERK, Cell Signaling Technology; catalog no.: 4370s, 1:1000 dilution), P38 (BOSTER Biological Technology Co, Ltd; catalog no.: BM4439, 1:1000 dilution), phospho-P38 (p-P38, Cell Signaling Technology; catalog no.: 4511s, 1:1000 dilution), Jun NH2-terminal kinase (JNK, BOSTER Biological Technology Co, Ltd; catalog no.: BM1219, 1:1000 dilution), phospho-JNK (p-JNK, Cell Signaling Technology, catalog no.: 9255s, 1:1000 dilution), β-actin (Santa Cruz; catalog no.: sc-47778, 1:1000 dilution), and GAPDH (MULTISCIENCES; catalog no.: Mab5465-100, 1:3000 dilution).

Techniques: shRNA, Knockdown, Activation Assay, Western Blot, Phospho-proteomics, Control, Infection

Journal: Materials Today Bio

Article Title: Mesenchymal stem cell-derived small extracellular vesicles-loaded GelMA microspheres enhance diabetic wound healing by promoting M2 macrophage polarization through p38 MAPK inhibition

doi: 10.1016/j.mtbio.2025.102423

Figure Lengend Snippet: MSC-sEVs promoted M2 polarization via inhibition of the p38 MAPK pathway. (A) Heatmap depicting cytokine levels in culture supernatants. (B) Heatmap illustrating the relative inflammatory cytokines levels and (C) Immune response-related genes in different treatment groups. (D) GSEA plots indicating enrichment of key pathway. (E) Venn diagram illustrating the overlap of genes across the pathways, with Mapk14 , Nfkb1 , and Fos identified as common regulators of inflammation and immune responses. (F) Bar graph showing the expression levels of key genes (n = 3). (G) Western blot analysis of p-MAPKAPK2, p-p38, and total p38 protein expression. (H–J) Quantification of (H) p38, (I) p-p38, and (J) p-MAPKAPK2 levels, demonstrating that MSC-sEVs treatment significantly reduces p-p38 and p-MAPKAPK2 levels. Data are presented as mean ± SD (n = 3). ∗ p < 0.05, ∗∗∗ p < 0.001.

Article Snippet: After blocking with 5 % BSA for 1 h at room temperature, membranes were incubated overnight at 4 °C with primary antibodies: CD9 (20597-1-AP, 1:3000, Proteintech, China), CD81 (27855-1-AP, 1:2000, Proteintech, China), Alix (12422-1-AP, 1:10000 Proteintech, China), calnexin (10427-2-AP, Proteintech, China), iNOS (22226-1-AP, 1:1000, Proteintech, China), CD206 (1:1000, 18704-1-AP, Proteintech, China), IL-6 (A0286,1:1000, ABclonal, China), IL-10 (A12255, 1:1000, ABclonal, China), p38 (A5049,1:1000, ABclonal, China), Phospho-p38 MAPK ( PAB43506 -P, 1:1000, Bioswamp, China), Phospho-MAPKAPK2 (AP0588, 1:1000, ABclonal, China) and GAPDH (10494-1-AP, 1:5000, Proteintech, China).

Techniques: Inhibition, Expressing, Western Blot