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: Cell

Article Title: Inflammation switches the chemoattractant requirements for naive lymphocyte entry into lymph nodes

doi: 10.1016/j.cell.2024.11.031

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Human CXCL12 (SDF-1a) Recombinant Protein , PeproTech , Cat# 300–28A.

Techniques: Control, Virus, Recombinant, Adjuvant, Reverse Transcription, RNAscope, SYBR Green Assay, Plasmid Preparation, Software