Journal: Nature communications

Article Title: Synovial microenvironment-influenced mast cells promote the progression of rheumatoid arthritis.

doi: 10.1038/s41467-023-44304-w

Figure Lengend Snippet: Fig. 1 | Frequency of synovial mast cells was increased in RA patients and correlated with disease severity. a Synovial membranes from OA and RA patients were digested into single-cell suspension. Synovial mast cells were gated as CD45+CD117+FcεRI+ cells. Numbers indicated the percentages of mast cells in the gates. b Statistical analysis of mast cell percentage in synovial single-cell suspen- sions and CD45+ immune cells (n = 10 biologically independent samples for each group, P = 0.0328 for percentage in suspensions, P = 0.0196 for percentage in CD45+ cells). c Synovial infiltrating mast cells were defined by tryptase immuno- fluorescence staining. Red, tryptase; Blue, DAPI. Scale bar: 75 μm. d Statistical analysis of mast cell numbers per high power field (HPF) in synovial sections (n = 10 biologically independent samples in OA group, n = 11 biologically independent samples in RA group, P = 0.0001). e Ki-67 expression on synovial mast cells was

Article Snippet: CD117-positive cells were isolated from synovial single-cell suspensions using a human CD117 microbead kit according to manufacturer’s instructions (Miltenyi Biotec, 130-091-332).

Techniques: Suspension, Staining, Expressing

Journal: Nature communications

Article Title: Synovial microenvironment-influenced mast cells promote the progression of rheumatoid arthritis.

doi: 10.1038/s41467-023-44304-w

Figure Lengend Snippet: Fig. 2 | RA synovial microenvironment-induced activation of mast cells. Syno- vial mast cells were enriched from digested synovial tissues of OA and RA patients by magnetic bead sorting for CD117+ cells and used for label-free mass spectro- metry. a Mast cell purity was confirmed by tryptase staining of enriched CD117+cells. Red, tryptase; Blue, DAPI. Scale bar: 25 μm. Data are representative of three independent experiments. b Heatmap of top 50 differentially expressed proteins between synovial mast cells from OA and RA patients by hierarchical clustering after z-score normalization. Upregulated and downregulated proteins in RA synovial mast cells were depicted in red and blue, respectively. c, d GO analysis and KEGG pathway analysis of differentially expressed proteins showed enriched terms including mitochondria metabolism, energy metabolism, and leukocyte

Article Snippet: CD117-positive cells were isolated from synovial single-cell suspensions using a human CD117 microbead kit according to manufacturer’s instructions (Miltenyi Biotec, 130-091-332).

Techniques: Activation Assay, Staining

Journal: International journal of molecular sciences

Article Title: Surgical Site-Released Tissue Is Potent to Generate Bone onto TCP and PCL-TCP Scaffolds In Vitro.

doi: 10.3390/ijms242115877

Figure Lengend Snippet: Figure 6. (a–c) ELISA boxplots of the osteogenic markers osteocalcin (a), osteopontin (b), and collagen I (c). Boxplots compare the concentration of ocn, opn, and col I in the control group with the PCL-TCP and β-TCP group over the cultivation time of 6 weeks. Groups are further subdivided into the condition’s agarose and glue, to examine if the chosen adhesive, used for the attachment of the scaffolds onto the plastic wells, makes a difference in protein expression. The boxblots indicating the median within the 25–75% percentile. The dots represent single outliners during measurements.

Article Snippet: Enzyme-linked immunosorbent assays (ELISAs) (human Duoset ELISA kit, R&D Systems, Bio Techne, Minneapolis, MN, USA) were then performed using the supernatants at a dilution of 1:10 for osteocalcin (1:50; pure for collagen and pure for osteopontin) according to the manufacturer’s protocol and measured at 450 nm using a Multiscan Ascent reader (Thermo Scientific).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Control, Adhesive, Expressing