Journal: Non-coding RNA Research

Article Title: Aerobic exercise alleviates allergic airway inflammation by suppressing circMETTL9 -mediated formation of macrophage extracellular traps ⋆

doi: 10.1016/j.ncrna.2025.08.008

Figure Lengend Snippet: Communication Between Airway Epithelial Cells and Macrophages Mediated by CXCL12-CXCR4 Regulates METs. (A) The co-expression network of IGFBP3 and macrophage chemokines was predicted by the STRING database. (B) The binding of CXCL12 to the CXCR4 receptor was predicted in the CellphoneDB database. (C) Representative SYTOX Green staining in macrophages treated with or without CXCL12 (n = 3). (D) The protein expression of MPO and CitH3 in macrophages was detected by Western blot analysis (n = 3). (E – G) ELISA was used to detect the expression of CXCL12 in the cell supernatant of the co-culture system (n = 3). RT-qPCR was used to detect the expression of CXCL12 in BEAS-2B cells and CXCR4 in macrophages (n = 3). All data are expressed as means ± SD. ∗ P < 0.05. GAPDH was used as a loading control for all Western blot assays. All data are expressed as means ± SD. ∗ P < 0.05.

Article Snippet: To assess the expression levels of CXCL12, ELISA kits specifically for mouse CXCL12 (catalog #KE10049, Proteintech, Chicago, USA), and for human CXCL12 ELISA Kit (catalog #RK00266, ABclonal, China) were utilized according to the manufacturer's protocols.

Techniques: Expressing, Binding Assay, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Quantitative RT-PCR, Control

Journal: Non-coding RNA Research

Article Title: Aerobic exercise alleviates allergic airway inflammation by suppressing circMETTL9 -mediated formation of macrophage extracellular traps ⋆

doi: 10.1016/j.ncrna.2025.08.008

Figure Lengend Snippet: Overexpression of CircMETTL9 Counteracts the Reduction Effect of Aerobic Exercise on METs. (A) Schematic timeline of the experimental protocol. Day 0: AAV-LUNG-OE- circMETTL9 by the nasal drip. Days 14, 28, 42, and 56 represent intraperitoneal (i.p.) injections of OVA. Days 35–68 represent exposure to ovalbumin aerosol. Aerobic exercise adaptation occurred from days 35–37, and days 39 and 67 represent the initial and final physical tests. Aerobic exercise was initiated on day 42 and ended on day 66. Euthanasia was performed on day 70. (B) The expression of circMETTL9 was detected by RNA FISH staining (n = 6). (C) The expression of circMETTL9 was performed by RT-qPCR (n = 6). (D) ELISA was used to detect the expression of CXCL12 in the BALF (n = 6). (E) The mRNA expression of CXCL12 and CXCR4 was detected by RT-qPCR (n = 6). (F) Western blot analysis was used to detect the protein expression of CitH3 and MPO in the lung tissue (n = 6). (G) Representative immunofluorescence images of CitH3, MPO, and CD68 staining of lung tissues. (H) The result of the Western blot showed the effect of circMETTL9 overexpression on IGFBP3 and EIF4A3 expression (n = 6). A, OVA-induced asthmatic mice and infected with blank AAV; E, mice were subjected to aerobic exercise and infected with blank AAV; OE-A, OVA-induced asthmatic mice and infected with AAV overexpressing circMETTL9 ; OE-AE, OVA-induced asthmatic mice performed aerobic exercise and infected with AAV overexpressing circMETTL9 . All data were shown as the means ± SDs and were assessed by a paired two-tailed t -test. ∗ P < 0.05.

Article Snippet: To assess the expression levels of CXCL12, ELISA kits specifically for mouse CXCL12 (catalog #KE10049, Proteintech, Chicago, USA), and for human CXCL12 ELISA Kit (catalog #RK00266, ABclonal, China) were utilized according to the manufacturer's protocols.

Techniques: Over Expression, Aerosol, Expressing, Staining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot, Immunofluorescence, Infection, Two Tailed Test