upper transwell chambers  (Corning Life Sciences)

Journal: Frontiers in Immunology

Article Title: Integrating scRNA-seq and machine learning identifies MNAT1 as a therapeutic target in OSCC

doi: 10.3389/fimmu.2025.1663487

Figure Lengend Snippet: Multi-cohort validation and functional experiment confirm MNAT1 as an oncogenic driver in OSCC. (A-C) MNAT1 expression is significantly upregulated in OSCC tumor tissues compared to normal tissues across three independent transcriptomic cohorts. (D) Kaplan-Meier analysis demonstrates reduced overall survival in OSCC patients with high MNAT1 expression versus low MNAT1 expression. (E) RT-qPCR confirms efficient MNAT1 knockdown in CAL27 cells. n = 3 biologically independent experiments. Data represent mean ± SD. P values were calculated by two-side Student’s t-test. (F, G) CCK-8 and colony formation assays reveal significantly impaired proliferative capacity in MNAT1-knockdown CAL27 cells. The quantitative analysis is shown on the right. n = 3 biologically independent experiments. Data represent mean ± SD. P values were calculated by two-side Student’s t-test. (H) Transwell migration and Matrigel invasion assays show markedly reduced migratory and invasive abilities following MNAT1 knockdown. Scale bar = 100um. The quantitative analysis is shown on the right. n = 3 biologically independent experiments. Data represent mean ± SD. P values were calculated by two-side Student’s t-test. (*p<0.05; **p<0.01; ***p<0.001; ****p<0.0001).

Article Snippet: For migration and invasion assays, resuspended in serum-free DMEM, 50,000 CAL27 cells were placed in Transwell upper chambers (NEST Biotechnology Co.,Ltd., China).

Techniques: Biomarker Discovery, Functional Assay, Expressing, Quantitative RT-PCR, Knockdown, CCK-8 Assay, Migration

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: THP-1 cells were incubated in serum-free medium for 12 h, then added to the upper Transwell chambers (Corning, NY, USA) at 5*10 4 /well.

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