mouse anti human bmp4  (R&D Systems)

Journal: Redox Biology

Article Title: Small molecule HCY-NBD stabilizes GSTM2 via cys174 sulfenylation to attenuate high glucose induced endothelial cell senescence and calcification

doi: 10.1016/j.redox.2026.104058

Figure Lengend Snippet: HCY-NBD inhibits high glucose-induced vascular calcification in vitro and in vivo (A-D) HUVECs were exposed to 0 or 5 μM HCY-NBD under either standard growth medium or DOM (30 mM glucose, 5 mM β -glycerophosphate, 50 μg/mL ascorbic acid) medium for 4 d, the protein levels of BMP2 and BMP4 were determined by Western blot, n = 3. (E – F) HUVECs were exposed to 0 or 5 μM HCY-NBD under either standard growth medium or DOM (30 mM glucose, 5 mM β -glycerophosphate, 50 μg/mL ascorbic acid) medium for 4 d, the protein levels of RUNX2 were determined by immunofluorescence, n = 3. (G – H) HUVECs were exposed to 0 or 5 μM HCY-NBD under either standard growth medium or DOM (30 mM glucose, 5 mM β -glycerophosphate, 50 μg/mL ascorbic acid) medium for 14 d, calcium deposition was evaluated by Alizarin Red S staining, n = 3. (I – J) En face immunofluorescence was performed on thoracic aortic segments from db/db mice to quantify RUNX2 expression in CD31-positive endothelial cells, using DAPI as nuclear counterstain, n = 4. (K) The activity of alkaline phosphatase (ALP) in mouse serum was measured, db/db: n = 4, other groups: n = 5. (L – M) Vascular calcification in thoracic aortas of db/db mice was evaluated using von Kossa staining, n = 3. One-way ANOVA followed by Tukey's post-hoc test was performed. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: GSTM2 ( AB196503 ), BMP4 ( AB124715 ), Ubiquitin ( AB137031 ), HA (66006-2-Ig) and Glutathione (AB19534) antibodies were sourced from Abcam. p21 (10355-1-AP), p16 (10883-1-AP), p53 (10442-1-AP), β -actin (66009-1-AP), BMP2 (66383-1-Ig), RUNX2 (20700-1-AP), GSTP1 (15902-1-AP), Flag (66008-1-Ig) and GFP (66002-1-Ig) antibodies were purchased from Proteintech.

Techniques: In Vitro, In Vivo, Western Blot, Immunofluorescence, Staining, Expressing, Activity Assay

Journal: Frontiers in Immunology

Article Title: Electroacupuncture alleviates pain by activating the MD2/TLR4/NF-κB pathway in the ST36 acupoint

doi: 10.3389/fimmu.2025.1626755

Figure Lengend Snippet: Functional enrichment analysis and distribution of differentially expressed proteins in the spinal cord of mice treated with EA and MD2 knockdown. (A) PCA analysis of all samples, each point denotes a sample, and different colors indicate different groups. (B) Each color represents a group, and the number of differentially expressed genes is indicated in the figure. (C) The hierarchical clustering results are presented as a dendrogram heatmap, with the x-axis and y-axis corresponding to samples and differentially expressed proteins, respectively. In the heatmap, red indicates significant upregulation, blue indicates significant downregulation, and gray represents the absence of quantitative data. (D, E) The vertical axis represents the GO level 2 functional annotation information, including molecular function, cellular component, and biological process. The horizontal axis indicates the significance of enrichment, with larger values on the horizontal axis representing a higher significance level of the corresponding GO functions. (F, G) KEGG pathway analysis of differential proteins. The vertical axis represents the names of pathways involving differentially expressed proteins. The horizontal axis indicates the significance of enrichment, with larger values signifying a higher level of significance for the enrichment in the corresponding pathway. The color gradient represents the magnitude of the enrichment factor (Rich Factor ≤ 1), with colors closer to red indicating a larger Rich Factor value. (H) Circos plot of the relationship between the significantly enriched KEGG pathway and the protein. (I) Bar chart showing the expression levels of the 5 key differentially expressed proteins. (J–L) The expression level of Grem1 (J) , BMP4 (K) , COX2 (L) (n = 3), ** P < 0.01 vs. the AIA+LVcon, # P < 0.01 vs. the AIA+LVcon+EA, ## P < 0.01 vs. the AIA+LVcon+EA.

Article Snippet: The blots were then washed and incubated overnight at 4°C with anti-MD2 antibody (1:1000, sc-80183, Santa, USA; 1:1000, ab24182, abcam, UK), anti-TLR4 antibody (1:1000, sc-293072, Santa, USA; 1:1000, 19811-1-AP, Proteintech Group, China), anti-P-P65 antibody (1:1000, 3033S, Cell Signaling Technology, USA), anti-P65 antibody (1:8000, 8242, Cell Signaling Technology, USA), anti-Grem1 antibody (1:800, sc-293426, Santa, USA), anti-BMP4 antibody (1:200, 12492-1-AP, Proteintech Group, Inc, USA), anti-COX2 antibody (1:500, A3560, ABclonal, China), anti-GAPDH antibody (1:3000000, A19056, ABclonal, China).

Techniques: Functional Assay, Knockdown, Expressing

Journal: Frontiers in Immunology

Article Title: Electroacupuncture alleviates pain by activating the MD2/TLR4/NF-κB pathway in the ST36 acupoint

doi: 10.3389/fimmu.2025.1626755

Figure Lengend Snippet: Functional enrichment analysis and distribution of differentially expressed proteins in the spinal cord of mice treated with EA and MD2 knockdown. (A) PCA analysis of all samples, each point denotes a sample, and different colors indicate different groups. (B) Each color represents a group, and the number of differentially expressed genes is indicated in the figure. (C) The hierarchical clustering results are presented as a dendrogram heatmap, with the x-axis and y-axis corresponding to samples and differentially expressed proteins, respectively. In the heatmap, red indicates significant upregulation, blue indicates significant downregulation, and gray represents the absence of quantitative data. (D, E) The vertical axis represents the GO level 2 functional annotation information, including molecular function, cellular component, and biological process. The horizontal axis indicates the significance of enrichment, with larger values on the horizontal axis representing a higher significance level of the corresponding GO functions. (F, G) KEGG pathway analysis of differential proteins. The vertical axis represents the names of pathways involving differentially expressed proteins. The horizontal axis indicates the significance of enrichment, with larger values signifying a higher level of significance for the enrichment in the corresponding pathway. The color gradient represents the magnitude of the enrichment factor (Rich Factor ≤ 1), with colors closer to red indicating a larger Rich Factor value. (H) Circos plot of the relationship between the significantly enriched KEGG pathway and the protein. (I) Bar chart showing the expression levels of the 5 key differentially expressed proteins. (J–L) The expression level of Grem1 (J) , BMP4 (K) , COX2 (L) (n = 3), ** P < 0.01 vs. the AIA+LVcon, # P < 0.01 vs. the AIA+LVcon+EA, ## P < 0.01 vs. the AIA+LVcon+EA.

Article Snippet: The blots were then washed and incubated overnight at 4°C with anti-MD2 antibody (1:1000, sc-80183, Santa, USA; 1:1000, ab24182, abcam, UK), anti-TLR4 antibody (1:1000, sc-293072, Santa, USA; 1:1000, 19811-1-AP, Proteintech Group, China), anti-P-P65 antibody (1:1000, 3033S, Cell Signaling Technology, USA), anti-P65 antibody (1:8000, 8242, Cell Signaling Technology, USA), anti-Grem1 antibody (1:800, sc-293426, Santa, USA), anti-BMP4 antibody (1:200, 12492-1-AP, Proteintech Group, Inc, USA), anti-COX2 antibody (1:500, A3560, ABclonal, China), anti-GAPDH antibody (1:3000000, A19056, ABclonal, China).

Techniques: Functional Assay, Knockdown, Expressing