mouse ccl2 je mcp 1  (R&D Systems)

Journal: Journal of Advanced Research

Article Title: Long noncoding RNA Nron and its functional motif modulate macrophage-mediated inflammatory response

doi: 10.1016/j.jare.2025.08.032

Figure Lengend Snippet: Nron deficiency in myeloid cells exacerbates systemic inflammatory responses in vivo . Nron fl/fl and Nron LysM-Cre mice were subjected to LPS- (10 mg/kg), or CLP-induced sepsis. A , BMDMs and PMs were isolated from Nron fl/fl and Nron LysM-Cre mice, and the levels of Nron were detected (n = 4). B and C , Kaplan–Meier survival analysis (n = 18 for LPS group, n = 22 for CLP group). D-J . LPS-induced sepsis (n = 6). Levels of inflammatory factors (IL-1β, IL-6, MCP-1 and TNF-α) in the serum (D) ; Representative images of lung biopsies stained with H&E, anti-F4/80 antibody or anti-Ly6G antibody (E) ; Quantitative analysis of lung injury (F) ; Quantitative analysis of F4/80 positive cells (G) ; Quantitative analysis of Ly6G positive cells (H) ; Western blot analysis for the phosphorylation and acetylation of p65 in lung tissues (I) ; Quantification of band density in panel I (J) . K-Q . CLP-induced sepsis (n = 6). Levels of inflammatory factors (IL-1β, IL-6, MCP-1 and TNF-α) in the serum (K) ; Representative images of lung biopsies stained with H&E, anti-F4/80 antibody or anti-Ly6G antibody (L) ; Quantitative analysis of lung injury (M) ; Quantitative analysis of F4/80 positive cells (N) ; Quantitative analysis of Ly6G positive cells (O) ; Western blot analysis for the phosphorylation and acetylation of p65 in lung tissues (P) ; Quantification of band density in panel P (Q) . Data are expressed as mean ± SEM. Statistical analysis was performed using Student's t -test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: ELISA kits were: tumor necrosis factor alpha (TNF-α) (RK00027, ABclonal), interleukin 6 (IL-6) (RK00008, ABclonal), IL-1β (RK00001, ABclonal), and monocyte chemoattractant protein-1 (MCP-1) (RK00381, ABclonal).

Techniques: In Vivo, Isolation, Staining, Western Blot, Phospho-proteomics

Journal: Journal of Advanced Research

Article Title: Long noncoding RNA Nron and its functional motif modulate macrophage-mediated inflammatory response

doi: 10.1016/j.jare.2025.08.032

Figure Lengend Snippet: MiR-146a-3p/miR-16-1-3p mediates the suppressive effect of Nron on SIRT1/NF-κB pathway. A , Venn diagram illustrating the overlap of miRNAs bound to Nron predicted by LncBase, lncRNASNP, and TargetScan databases. B , Relative luciferase activities of luciferase reporters containing Nron-WT or Nron-MUT in RAW246.7 cells treated with candidate miRNA mimics (n = 6 for WT group; n = 4 for MUT group). C and D , BMDMs challenged with LPS were treated with miR-146a-3p mimic, and the phosphorylation/acetylation of p65 (C) and the expression of Il-6 and Mcp-1 (D) were determined (n = 3). E and F , BMDMs challenged with LPS were treated with miR-16-1-3p mimic, and the phosphorylation/acetylation of p65 (E) and the expression of Il-6 and Mcp-1 (F) were determined (n = 3). G , BMDMs challenged with LPS were treated with miR-744-3p mimic, and the phosphorylation/acetylation of p65 were determined (n = 3). H , The protein levels of SIRT1 in BMDMs treated with miR-146a-3p, miR-16-1-3p or miR-744-3p mimic for 24 h were determined (n = 5). I and J , Relative luciferase activities of luciferase reporters containing WT or MUT Sirt1 3′-UTR in RAW246.7 cells treated with miR-146a-3p (I) or miR-16-1-3p (J) mimic (n = 6). K, RIP assay was performed using anti-Ago2 or IgG antibodies, and the relative enrichment of Nron, miR-146a-3p and miR-16-1-3p in immunoprecipitated RNA was determined by qRT-PCR (n = 3). L, upper panel : The expression of miR-146a-3p and miR-16-1-3p was determined in BMDMs infected with Ad-Nron or Ad-GFP; lower panel : The expression of Nron was determined in BMDMs treated with miR-146a-3p or miR-16-1-3p mimic (n = 3). M, WT and NKO BMDMs challenged with LPS were treated with anta-miR-146a-3p or anta-miR-16-1-3p, and the phosphorylation/acetylation of p65 were determined (n = 3). N, WT and NKO BMDMs were treated with anta-miR-146a-3p or anta-miR-16-1-3p for 24 h, and the protein levels of SIRT1 were determined (n = 4). Data are expressed as mean ± SEM. Statistical analysis was performed using Student's t -test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: ELISA kits were: tumor necrosis factor alpha (TNF-α) (RK00027, ABclonal), interleukin 6 (IL-6) (RK00008, ABclonal), IL-1β (RK00001, ABclonal), and monocyte chemoattractant protein-1 (MCP-1) (RK00381, ABclonal).

Techniques: Luciferase, Phospho-proteomics, Expressing, Immunoprecipitation, Quantitative RT-PCR, Infection

Journal: Journal of Advanced Research

Article Title: Long noncoding RNA Nron and its functional motif modulate macrophage-mediated inflammatory response

doi: 10.1016/j.jare.2025.08.032

Figure Lengend Snippet: The functional motif of Nron regulates macrophage inflammatory responses. A and B, BMDMs challenged with LPS were infected with Ad-NCM1 (A) or Ad-NCM2 (B) , and the phosphorylation/acetylation of p65 were determined (n = 3). C, BMDMs challenged with LPS were infected with Ad-NCM2, and the expression of Il-β , Il-6 , Mcp-1 , Il-12a and Il-23 were determined (n = 3). D, BMDMs challenged with LPS were infected with Ad-Nron or Ad-NCM2, and the phosphorylation/acetylation of p65 were determined (n = 3). E and F, Human THP1 cells challenged with LPS were infected with Ad-NCM2 (h), and the phosphorylation/acetylation of P65 (E) and the expression of IL-β , IL-6 , MCP-1 , IL-12a and IL-23 (F) were determined (n = 3). G, The expression of miR-146a-3p and miR-16-1-3p was determined in BMDMs infected with Ad-NCM2 or Ad-GFP (n = 3). H, BMDMs with NCM2 overexpression were treated with miR-16-1-3p mimic for 24 h, and the protein levels of SIRT1 were determined (n = 4). I-Q, Sirt1 fl/fl and Sirt1 LysM-Cre mice were treated with AAV6-NCM2 by intravenous/intratracheal administration, and then subjected to LPS- (10 mg/kg)-induced sepsis (n = 15 for Sirt1 fl/fl group, n = 18 for Sirt1 LysM-Cre group in survival analysis; n = 6 for other assays). I, The expression efficiency of AAV-NCM2 in the alveolar macrophages. J, Kaplan–Meier survival analysis. K, The levels of inflammatory factors (IL-1β, IL-6 and MCP-1) in the serum. L, Representative images of lung biopsies stained with H&E, anti-F4/80 antibody or anti-Ly6G antibody. M, Quantitative analysis of lung injury. N, Quantitative analysis of F4/80 positive cells. O, Quantitative analysis of Ly6G positive cells. P, Western blot analysis for the phosphorylation/acetylation of p65 and SIRT1 in alveolar macrophages. Q, Quantification of band density in panel P . Data are expressed as mean ± SEM. Statistical analysis was performed using Student's t -test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: ELISA kits were: tumor necrosis factor alpha (TNF-α) (RK00027, ABclonal), interleukin 6 (IL-6) (RK00008, ABclonal), IL-1β (RK00001, ABclonal), and monocyte chemoattractant protein-1 (MCP-1) (RK00381, ABclonal).

Techniques: Functional Assay, Infection, Phospho-proteomics, Expressing, Over Expression, Staining, Western Blot