immunofluorescent staining cenp f d6x4l rabbit mab cell signaling technology 58982t 250 immunofluorescent staining donkey anti mouse igg h l  (Cell Signaling Technology Inc)

Journal: Advanced Healthcare Materials

Article Title: Structured Polymers Enable the Sustained Delivery of Glucocorticoids within the Intra‐Articular Space

doi: 10.1002/adhm.202403000

Figure Lengend Snippet: All groups consisted of intra‐articular injection with 10 µL of either saline control (CON), blank GSH (GEL: 488 1.5% gellan (w/v), 10 mM NaCl), methylprednisolone sodium succinate (PRED: 10 mg mL −1 ) or GSH loaded with methylprednisolone sodium succinate (PRED GEL: Gel: 1.5% gellan (w/v), 10 mM NaCl, Pred 10 mg mL −1 ). A) Representative image of Coomassie blue dye loaded gellan sheared hydrogel (GSH) administered into the intra‐articular space in the TNFtg murine model of polyarthritis. B) Body weight, C) disease score, and D) Inflammatory paw score D) measured over 21 days in the TNFtg model of polyarthritis following intra‐articular injection. E) Representative sagittal images of formalin‐fixed paraffin embedded knee joints, stained with hematoxylin and eosin, and F) measurement of synovitis, G) sub‐chondral pannus formation, and H) surface cartilage area in TNFtg animals at day 21 following treatment. I‐K) Fluorescence intensity/um 2 of synovial tissue as a measure of expression of TNFα and podoplanin, and representative images of fluorescent staining for RANKL, TNFa, podoplanin, and DAPI, determined by confocal immunofluorescence area in TNFtg animals at day 21 following treatment. L) Representative GILZ staining within synoviocytes was determined in the same groups within articular synovium by immunohistochemistry. M) Representative alcian blue staining within the articular space was performed in TNFtg animals after 21 days following injection with either saline control (Con) or GSH loaded with methylprednisolone sodium succinate at 20 and 60X magnification. Black arrows denote positively stained GSHs. Data are presented as mean ± SEM of at least three six animals per group. Statistical significance was determined using one‐way ANOVA with Tukey's multiple comparisons test (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). (scale bars, 50 µm).

Article Snippet: The following primary antibodies were used for immunofluorescence staining of tissues: goat anti‐mouse podoplanin (R&D systems, AF3244,1 in 100), mouse anti‐mouse RANKL (Proteintech, Cat No: 66610‐1‐Ig, 1 in 100), and rabbit ant‐mouse TNF‐α (Proteintech, Cat. No: 80258‐6‐RR, 1 in 50).

Techniques: Injection, Saline, Control, Formalin-fixed Paraffin-Embedded, Staining, Fluorescence, Expressing, Immunofluorescence, Immunohistochemistry

Journal: Biomaterials

Article Title: Integrated-omics profiling unveils the disparities of host defense to ECM scaffolds during wound healing in aged individuals.

doi: 10.1016/j.biomaterials.2024.122685

Figure Lengend Snippet: Fig. 2. The acute inflammation rather than type II immune response around ECM scaffold membrane in aged wounds. (a) Schematic diagram for generating the scRNA-seq and spatial transcriptomics data from ECM mediated wound healing at POD7 in young and aged mice. (b) The UMAP reduction results reveals 13 different cell types in wounds. The marker genes are listed on the right of the annotation. (c) The annotated clustering results for spatial transcriptomic based on the anatomic structure of the wounds. (d) The proportion of each cell type of the whole wounds and their related wound healing process. (e) The upregulated results for Gene Sets Enrichment Analysis (GSEA) in ECM scaffolds treated aged wounds. (f) The AddModule scores of acute inflammation term in spatial transcriptomics expression. (g) The IF results for INOS and LY6G in Young_ECM and Aged_ECM at POD7. (Scale bars at the same column are the same) (h–i) The semiquantitative analysis of INOS and LY6G fluorescence area in (g) respectively. **P < 0.01 and *P < 0.05 by Student’s t-test (n = 5) for data in (h) and (i).

Article Snippet: For the evaluation the acute inflammatory molecules and immune cells infiltration, immunohistochemistry staining for CD3 (14-0032-82, Thermo Fisher Scientific, 1:100) and immunofluorescent staining for LY6G (65140-1-Ig, Proteintech, 1:200), INOS (22226-1-AP, Proteintech, 1:200), IL-1β (P420B, Thermo Fisher Scientific, 1:200),TNF-α (60291-1-Ig Proteintech, 1:200), OPN (22952- 1-AP, Proteintech, 1:200), F4/80 (29414-1-AP, Proteintech, 1:200), ARG1 (16001-1-AP, Proteintech, 1:150), IL-10 (ARC9102, Thermo Fisher Scientific, 1:100), CD62L (26477-1-AP Proteintech, 1:200), CD4 (67786-1-Ig, Proteintech, 1:500) were performed.

Techniques: Membrane, Marker, Expressing, Fluorescence