anti il8  (R&D Systems)

Journal: Nature Communications

Article Title: HMGA1 orchestrates chromatin compartmentalization and sequesters genes into 3D networks coordinating senescence heterogeneity

doi: 10.1038/s41467-024-51153-8

Figure Lengend Snippet: a Single-cell UMAP projection of Grow, OIS and OIS-shA1 cells (coloured by condition but each representing two replicates), highlighting key senescence genes: MKI67 , IL8 , CXCL1 , IL1B , MMP3 , and CDKN1A ; expression values (log-transformed) were scaled to be between 0 and 10 for visualisation, with grey representing no expression detected. b Distribution of UCell single-cell scores for selected MSigDB Hallmarks gene sets in n = 6165 Grow, n = 2828 OIS, and n = 2073 OIS-shA1 cells; P-values derived from two-sided Wilcoxon testing. Box plot centre line represents the median, the bounds correspond to the 0.25 and 0.75 quantiles, the whiskers represent the 0.1 and 0.9 quantiles. c UMAP projection of the OIS and OIS-shA1 conditions only for Milo testing of cell neighbourhoods and clustering based on the log-fold changes between OIS and OIS-shA1. d Representative markers of the four Milo clusters of differential expression between OIS and OIS-shA1 cells at single-cell level, with expression values scaled between 0 and 1 and averaged over the cell neighbourhoods in each cluster from c . e Representative markers for clusters 1–4 coloured by scaled expression values on the UMAP projection of OIS and OIS-shA1 cells. f Schematic representation of the features of the four clusters of senescent cells identified using overrepresentation analysis, highlighting the clusters over-represented in OIS (2 and 4) and in OIS-shA1 (1 and 3), respectively. Although inflammatory SASP (iSASP) and p16 have been collectively considered senescence hallmarks, they represent distinct types of senescence at the single-cell level. Clusters 3 and 4 express cell-cycle genes and Cluster 4 resembles the previously described NOTCH-related ‘early phase senescence’ with augmented fibroblastic features , . g Dimensionality reduction (PCA) of the log-fold changes of OIS cells (bulk RNA-seq) in response to shHMGA1, shCEBPB, shp53 and double knock-down of p53 and CEBPB . h UMAP projection of the Grow and OIS cells, coloured by UCell scoring of the gene signatures of the genes activated in OIS and up-regulated by shA1 (repressed by HMGA1, top) and down-regulated by the double knock-down of p53 and CEBPB (activated by p53 + CEBPB , bottom).

Article Snippet: The following primary antibodies were used for immunofluorescence: anti-H3K9me3 (mouse monoclonal, Hiroshi Kimura Laboratory, clone CMA318, 4 μg/mL), anti-HMGA1 (rabbit polyclonal, Cold Spring Harbor Labs, CS2637, 1:1000), anti-CDKN2A p16 (mouse monoclonal, BD Biosciences 554079, clone G175-1239, 1:500), and anti-IL8 (mouse monoclonal, R&D Systems, MAB208, 1:500).

Techniques: Expressing, Transformation Assay, Derivative Assay, RNA Sequencing Assay, Knockdown

Journal: International immunopharmacology

Article Title: Oxidative stress induces ferroptosis in tendon stem cells by regulating mitophagy through cGAS-STING pathway.

doi: 10.1016/j.intimp.2024.112652

Figure Lengend Snippet: Fig. 4. In TSCs, H2O2 sensitized the cGAS-STING axis⋅H2O2 at a concentration of 100 μM was applied to TSCs for 24 h. (A) Western blot was used to assess the protein levels of the cGAS-STING axis. (B) Immunofluorescence was used to figure out how cGAS and STING were expressed. (C-E) RT-PCR was used to detect the mRNA expression of particular gene, including cGAS, STING, IFNβ, IL-1β, IL-6 and IL-8. (F) The protein levels of proinflammatory factors downstream of the cGAS-STING axis were detected by Western blot, including IL-1β, IL-6, IL-8. (n = 3); *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 #p < 0.05, ##p < 0.01, ###p < 0.001, &&p < 0.01, &&&p < 0.001.

Article Snippet: Anti-IL8 antibody (ab207612) was apply for trial installation from Santa Cruz Biotechnology.

Techniques: Concentration Assay, Western Blot, Immunofluorescence, Reverse Transcription Polymerase Chain Reaction, Expressing