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

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: CXCL12/CXCR4 modulates macrophage efferocytosis to induce glomerular crescent formation and fibrosis via ELMO1/DOCK180/RAC1 signaling in ANCA-associated glomerulonephritis

doi: 10.1007/s00018-025-05750-5

Figure Lengend Snippet: Release of the chemokine CXCL12 by endothelial cells in AAGN promoted monocyte recruitment and infiltration. A Spatial transcriptome sequencing of renal tissues revealed that CCL2, CCL21 and CXCL12 were increased in AAGN group, of which CXCL12 was the highest. B The immunohistochemical staining of CXCL12 was increased in AAGN group (AAGN = 6, CTRL = 4, 80 ×, scale bar = 25 μm). C ELISA levels of CXCL12 were elevated in MPO + AAGN plasma ( n = 15) and urine ( n = 6). D ELISA levels of CXCL12 were not significantly increased in PR3 + AVV plasma ( n = 10) and PR3 + AAGN urine ( n = 9). E CXCL12 mRNA was increased in EA.hy926 after 3 days of incubation with AAGN serum medium, but not in HRMC, HPC and HK-2 ( n = 3). F Western blotting showed that after 3 days of incubation with AAGN serum medium, CXCL12 protein level was increased in EA.hy926, while not changed in HRMC, HPC and HK-2 ( n = 3). G CD14 immunofluorescence was increased in the crescents of AAGN group (AAGN = 6, CTRL = 4, PAS 80 ×, scale bar = 25 μm, IF 600 ×, scale bar = 100 μm). H Transwell assay confirmed that EA.hy926 incubated with AAGN serum in the lower chamber could recruit CD14. + monocytes from the upper chamber to migrate to the lower chamber ( n = 3, 400 ×, scale bar = 10 μm). * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: CXCL12 (CSB-EQ027490HU for human, CSB-E08729r for rat, Cusabio, Wuhan, China) and soluble CXCR4 (CSB- E12825 h for human, CSB-E12703r for rat, Cusabio) in serum and urine were measured with ELISA kits following the manufacturer’s instructions.

Techniques: Sequencing, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Incubation, Western Blot, Immunofluorescence, Transwell Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: CXCL12/CXCR4 modulates macrophage efferocytosis to induce glomerular crescent formation and fibrosis via ELMO1/DOCK180/RAC1 signaling in ANCA-associated glomerulonephritis

doi: 10.1007/s00018-025-05750-5

Figure Lengend Snippet: Glomerular endothelial cell apoptosis in AAGN resulted in CXCL12 release. A Immunofluorescence levels of CD31 and ERG were decreased in AAGN crescents, whereas TUNEL was increased (AAGN = 6, CTRL = 4, PAS 80 ×, scale bar = 50 μm, IF 600 ×, scale bar = 20 μm). B The mRNA and protein levels of BCL-2/BAX were decreased in endothelial cells incubated with AAGN serum ( n = 3). C The mRNA and protein levels of cleaved-Caspase-3 were decreased in endothelial cells incubated with AAGN serum ( n = 3). D The mRNA and protein levels of CD31 and ERG were decreased in endothelial cells incubated with AAGN serum ( n = 3). E CCK-8 assay showed that with the prolonged incubation time of AAGN serum, the proliferation ability of endothelial cells decreased, and the most obvious was observed at 72–84h ( n = 3). F Flow cytometry verified that the proportion of apoptotic endothelial cells was significantly increased when incubated with AAGN serum for 3 days ( n = 3). * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: CXCL12 (CSB-EQ027490HU for human, CSB-E08729r for rat, Cusabio, Wuhan, China) and soluble CXCR4 (CSB- E12825 h for human, CSB-E12703r for rat, Cusabio) in serum and urine were measured with ELISA kits following the manufacturer’s instructions.

Techniques: Immunofluorescence, TUNEL Assay, Incubation, CCK-8 Assay, Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: CXCL12/CXCR4 modulates macrophage efferocytosis to induce glomerular crescent formation and fibrosis via ELMO1/DOCK180/RAC1 signaling in ANCA-associated glomerulonephritis

doi: 10.1007/s00018-025-05750-5

Figure Lengend Snippet: Inhibition of CXCL12/CXCR4 alleviates AAGN progression. A Suitable concentrations for LIT927 (CXCL12 neutral ligand antagonist) and AMD3100 (CXCR4 inhibitor) were determined as 30 μM and 100 μM, respectively, using CCK-8 assays ( n = 3). B When mixed individually with AAGN serum to stimulate macrophages (Mφs), both drugs reduced MERTK, CD163, TGF-β1 and ELMO1/DOCK180/RAC1 axis components at transcriptional and protein levels ( n = 3). C Schematic of the EAV rat model establishment. Drug administration, either concurrently with modeling or from the third week post-modeling, did not significantly affect rat body weight D but alleviated hematuria E and proteinuria F , with AMD3100 showing greater efficacy. Starting treatment at the third week was as effective as concurrent administration. G LIT927 administration at the onset of modeling reduced cellular and fibrous crescent formation. When started from the third week, it alleviated fibrocellular and fibrous crescents without affecting cellular crescents. AMD3100 reduced various types of crescent formation when given at modeling onset but primarily alleviated fibrocellular and fibrous crescents when started three weeks later (80 ×, scale bar = 10 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001

Article Snippet: CXCL12 (CSB-EQ027490HU for human, CSB-E08729r for rat, Cusabio, Wuhan, China) and soluble CXCR4 (CSB- E12825 h for human, CSB-E12703r for rat, Cusabio) in serum and urine were measured with ELISA kits following the manufacturer’s instructions.

Techniques: Inhibition, CCK-8 Assay

Journal: International Journal of Molecular Sciences

Article Title: Derivation of Human Corneal Keratocytes from ReLEx SMILE Lenticules for Cell Therapy and Tissue Engineering

doi: 10.3390/ijms24108828

Figure Lengend Snippet: hCSC characteristics: ( A ) percentage of various fibroblast and keratocyte phenotype markers, adhesive molecule markers in two hCSC populations: CFs ( n = 13), rCFs ( n = 8); * p < 0.05, ** p = 0.003, # p = 0.01. ( B ) Percentage of CFs and rCFs that expressed keratocyte markers. ( C ) Fluorescence microscopy images showing the comparative expression of various markers in hCSCs: green—stained with fibronectin and α-smooth muscle α actin antibodies (induction of myofibroblasts with addition of TGFβ1, positive control), blue—stained with DAPI for nuclei. The scale bar is 100 µm for the fibronectin and 20 µm for the other markers.

Article Snippet: Commercially available ELISA kits were used to determine the human insulin growth factor-1 (IGF-1), human free brain-derived neurotrophic factor (BDNF) (all kits: R&D Systems, Minnesota, MN, USA), human stromal-derived factor-1a (SDF-1a; Thermo Scientific, Waltham, MA, USA), human vimentin, total collagen, human fibronectin (all kits: Abcam, Boston, MA, USA), human soluble intercellular adhesion molecule-1 (sICAM-1; Affymetrix eBioscience, Santa Clara, CA, USA), vascular endothelial growth factor (VEGF), erythropoietin (EPO), and tumor necrosis factor-alpha (TNF-α) (all kits: Vector-BEST, Novosibirsk, Russia), produced by the CFs and rCFs.

Techniques: Fluorescence, Microscopy, Expressing, Staining, Positive Control