Journal: Journal of experimental & clinical cancer research : CR

Article Title: Gastric cancer-derived exosomal let-7 g-5p mediated by SERPINE1 promotes macrophage M2 polarization and gastric cancer progression.

doi: 10.1186/s13046-024-03269-4

Figure Lengend Snippet: Fig. 2 High SERPINE1 expression in GC cells promotes macrophage M2 polarization. tSNE visualization of nine single-cell clusters partitioned by unsu pervised cluster analysis, SERPINE1 expression of each single-cell, and SERPINE1 expression abundance of different single-cell clusters in the GSE134520 (A–C) and GSE167297 (D–F) datasets. (G) Flow cytometry analysis of the proportion of CD68+CD206+ macrophages in a Transwell co-culture system, with MKN45 and AGS cells overexpressing (oe_SERPINE1) or silencing SERPINE1 (shRNA#3 or sh_SERPINE1#3) in the upper chamber, and THP1 cells treated with PMA in the lower chamber. (H) Immunofluorescence staining of xenograft tumor tissues. Comparison of the proportion of M1 or M2 macrophage infiltra tion. Green indicates F4/80. Red indicates iNOS or Arg1 expression

Article Snippet: THP-1 cells were differentiated into macrophages using 150 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma) for 24 h and subsequently co-cultured with cancer-derived exosomes or GC cells in 6-well plates with 0.4-μm membranes for 72 h. Harvested macrophages were converted into single-cell suspensions, stained with Elab Fluor 488 anti-human CD68 (Mouse, 1:20, ElabScience) and APC anti-human CD206 (Mouse, 1:20, ElabScience) antibodies, and analyzed for CD68+CD206+ populations by flow cytometry (Accuri C6, BD).

Techniques: Expressing, Flow Cytometry, Co-Culture Assay, shRNA, Immunofluorescence, Staining, Comparison

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Gastric cancer-derived exosomal let-7 g-5p mediated by SERPINE1 promotes macrophage M2 polarization and gastric cancer progression.

doi: 10.1186/s13046-024-03269-4

Figure Lengend Snippet: Fig. 5 SERPINE1-mediated gastric cancer-derived exosomes facilitate the polarization of THP1 cells into M2 macrophages. (A) Schematic representation of the extraction and identification of exosomes and the induction of macrophage polarization. Transmission electron microscopy (B), nanoparticle tracking analysis (C), and western blotting (D) were used to identify the morphology, particle size, and markers of exosomes. (E) Confocal laser scanning microscopy detected Dil-labeled exosomes (red) internalized by DAPI-labeled macrophages (blue). (F–G) Immunofluorescence analysis of the proportion of CD206+ cells in THP1 cells treated with exosomes. (H–I) Flow cytometry analysis of the proportion of CD68+CD206+ cells in THP1 cells treated with exosomes. (J–K) qRT-PCR analysis of M1 markers (iNOS and TNF-α) and M2 markers (TGF-β, IL-10, and Arg-1) in THP1 cells treated with exosomes. (L–N) Transwell migration and invasion assays of GC cells (upper chamber) co-cultured with macrophages (lower chamber) ingesting exosomes

Article Snippet: THP-1 cells were differentiated into macrophages using 150 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma) for 24 h and subsequently co-cultured with cancer-derived exosomes or GC cells in 6-well plates with 0.4-μm membranes for 72 h. Harvested macrophages were converted into single-cell suspensions, stained with Elab Fluor 488 anti-human CD68 (Mouse, 1:20, ElabScience) and APC anti-human CD206 (Mouse, 1:20, ElabScience) antibodies, and analyzed for CD68+CD206+ populations by flow cytometry (Accuri C6, BD).

Techniques: Derivative Assay, Extraction, Transmission Assay, Electron Microscopy, Western Blot, Confocal Laser Scanning Microscopy, Labeling, Immunofluorescence, Flow Cytometry, Quantitative RT-PCR, Migration, Cell Culture

Journal: Advanced Science

Article Title: Synovium‐On‐A‐Chip: Simulating the Microenvironment of the Rheumatoid Arthritis Synovium via Multicell Interactions to Target Fibroblast‐Like Synoviocytes

doi: 10.1002/advs.202511945

Figure Lengend Snippet: Expression levels of inflammatory factors in cocultures of RA FLSs, M1 macrophages, and HUVECs at different seeding ratios. A) Heatmap illustrating the expression levels of multiple inflammation‐related factors secreted by RA FLSs cultured alone (F), RA FLSs cocultured with M1 macrophages (FM), and RA FLSs, M1 macrophages, and HUVECs cocultured at ratios of 1:1:2 (FME‐2), 1:1:1 (FME‐1), and 1:1:0.5 (FME‐0.5). B) Expression levels of interleukin (IL)‐1β, IL‐6, IL‐8, and tumor necrosis factor‐alpha (TNF‐α) in the F, FM, FME‐2, FME‐1, and FME‐0.5 groups. C) Expression levels of granulocyte macrophage‐colony stimulating factor (GM‐CSF), C‐X‐C motif chemokine ligand 10 (CXCL10), IL‐12p70, interferon (IFN)‐γ, IFN‐α, IFN‐β, IFN‐λ1, and IFN‐λ2 in the F, FM, FME‐2, FME‐1, and FME‐0.5 groups. D) Multiplex immunofluorescence staining for specific proteins, including Vimentin (mesenchymal marker), CD68 (macrophage marker), and CD31 (endothelial cell marker), in RA synovial tissue. Scale bars, 100 µm. n = 3 per group. Data are presented as the mean ± SD. Statistical analysis: one‐way ANOVA with Tukey's test. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: For flow cytometry, primary cells were fixed, permeabilized, and subsequently incubated with Alexa Fluor 488‐conjugated anti‐Vimentin (677809; BioLegend, USA) and Elab Fluor 647 Anti‐Human CD68 (E‐AB‐F1299M; Elabscience, China) antibodies.

Techniques: Expressing, Cell Culture, Multiplex Assay, Immunofluorescence, Staining, Marker