Journal: Science Advances

Article Title: Pro-inflammatory megakaryocyte gene expression in murine models of breast cancer

doi: 10.1126/sciadv.abo5224

Figure Lengend Snippet: CD41 + DAPI low cells were isolated from the BM of a 13-week-old PyMT mouse and a WT littermate for scRNA-seq. ( A ) Dimensionality reduction using Uniform Manifold Approximation and Projection (UMAP) of pooled PyMT and WT samples. Cells of MK lineage (orange) were separated into five clusters (G 1 = 1699, G 2 /M = 1395, MK1 = 871, MK2 = 537, and MK3 = 344 cells) based on differential gene expression. Gray cells from non-MK clusters ( n = 4184 cells) were excluded. ( B ) Overlay of PyMT (red, n = 3817) and WT (blue, n = 5213) cells. ( C ) Heatmaps clustered according to average log fold change depicting the top 50 differentially expressed genes between PyMT and WT cells in MK1, MK2, and MK3. Scatterplots depict gene expression of all recorded transcripts. Red lines model no change in gene expression between WT and PyMT samples. ( D ) Diseases and functions associated with differentially expressed genes in MKs from PyMT mice. ( E ) Expression levels of thrombospondin 1 ( Thbs1 ), cathepsin G ( Ctsg ), S100a8 , and S100a9 mRNA transcripts among MK subclusters.

Article Snippet: In some cases, tumor cells and platelet releasate were pretreated with the S100A8/A9 inhibitor paquinimod (Sigma-Aldrich, SML2883) for 30 min before the addition of tumor cells to transwells.

Techniques: Isolation, Expressing

Journal: Science Advances

Article Title: Pro-inflammatory megakaryocyte gene expression in murine models of breast cancer

doi: 10.1126/sciadv.abo5224

Figure Lengend Snippet: ( A ) Heatmap of differentially expressed ( P < 0.01) proteins from native BM MKs of mice bearing Met-1 tumors for 21 days or tumor-naïve controls (PBS); n = 5 mice per condition. ( B ) Diseases and functions of differentially expressed proteins from Met-1 and PBS-only injected mice. The number of proteins associated with each disease or function is shown to the right of the plot. ( C ) Volcano plot highlighting differentially expressed proteins. Factors of interest are annotated. ( D ) Protein expression of candidate proteins S100A8 (S100a8), lipocalin-2 (Lcn2), and cathepsin G (Ctsg). n = 5 mice per condition. P values were determined by two-sample t tests. ( E ) S100A8/A9 levels in MKs isolated from 13-week-old PyMT and WT mice as determined by ELISA. n = 5 mice per condition. P value determined by two-sample t test.

Article Snippet: In some cases, tumor cells and platelet releasate were pretreated with the S100A8/A9 inhibitor paquinimod (Sigma-Aldrich, SML2883) for 30 min before the addition of tumor cells to transwells.

Techniques: Injection, Expressing, Isolation, Enzyme-linked Immunosorbent Assay

Journal: Science Advances

Article Title: Pro-inflammatory megakaryocyte gene expression in murine models of breast cancer

doi: 10.1126/sciadv.abo5224

Figure Lengend Snippet: ( A ) Platelet counts, mean platelet volume (MPV), platelet distribution width (PDW), and immature platelet fraction in 7- to 13-week-old PyMT and WT mice. n = 10 mice per condition. * P < 0.05 and ** P < 0.01, as determined by one-way ANOVA. ( B ) Thiazole orange (TO) staining of platelets from PyMT and WT mice. n = 6 to 7 mice per condition. P values were determined by two-sample t tests. ( C ) Proportion of biotin + platelets isolated from PyMT and WT mice following in vivo labeling with biotin (0 hours). Dashed line depicts basal signal from unlabeled platelets. n = 5 mice per condition. Error bars depict SEM. ( D ) Plasma levels of platelet factor 4 (PF4) from PyMT and WT mice. n = 9 to 11 mice per condition. P values were determined by two-sample t tests. ( E ) Surface expression of P-selectin (CD62p) on the surface of platelets isolated from PyMT or WT mice following thrombin stimulation. n = 4 to 5 mice per condition. Error bars depict SEM. ( F ) S100A8 and S100A8/A9 levels in platelets from PyMT and WT mice. n = 9 to 19 mice per condition. P values were determined by two-sample t tests. ( G ) S100A8/A9 from the plasma of PyMT and WT mice. n = 4 to 7 mice per condition. ( H ) Lcn-2 levels of platelets from 13-week-old PyMT and WT mice. n = 5 mice per condition. P values were determined by two-sample t tests. ( I ) Lcn-2 levels from the plasma of PyMT and WT mice. n = 3 mice per condition. P values were determined by two-sample t tests.

Article Snippet: In some cases, tumor cells and platelet releasate were pretreated with the S100A8/A9 inhibitor paquinimod (Sigma-Aldrich, SML2883) for 30 min before the addition of tumor cells to transwells.

Techniques: Staining, Isolation, In Vivo, Labeling, Expressing

Journal: Science Advances

Article Title: Pro-inflammatory megakaryocyte gene expression in murine models of breast cancer

doi: 10.1126/sciadv.abo5224

Figure Lengend Snippet: ( A ) Median age, platelet count, and MPV from recently diagnosed and untreated patients with early-stage (I to III) breast cancer. ( B ) Plasma levels of PF4 from breast cancer patients and age-matched cancer-free female controls. n = 6 control subjects and 9 patients per condition. ( C ) Levels of S100A8/A9 in platelets isolated from cancer-free subjects and early-stage breast cancer patients. n = 6 to 10 subjects/patients per condition. ( D ) S100A8/A9 levels in the plasma and serum of cancer-free subjects and early-stage breast cancer patients. n = 5 to 10 subjects/patients per condition. N/A in cancer patients indicates no assigned stage due to the lack of lymph node examination. P values were determined by one-way ANOVA.

Article Snippet: In some cases, tumor cells and platelet releasate were pretreated with the S100A8/A9 inhibitor paquinimod (Sigma-Aldrich, SML2883) for 30 min before the addition of tumor cells to transwells.

Techniques: Isolation

Journal: Frontiers in Pharmacology

Article Title: Shuangxinfang Prevents S100A9-Induced Macrophage/Microglial Inflammation to Improve Cardiac Function and Depression-Like Behavior in Rats After Acute Myocardial Infarction

doi: 10.3389/fphar.2022.832590

Figure Lengend Snippet: The effect of PCF on the S100A9/NF-κB pathway in the hippocampus and myocardium. (A) The gene expression of S100A9 and NF-κB in the hippocampus and myocardium detected by RT-qPCR. Data were presented as mean ± SD, n = 3 per group. (B) Immunofluorescence analysis of S100A9 in hippocampus tissues. S100A9 immunostaining was shown in green and DAPI in blue. (C) Mean optical density of S100A9-positive cells. Three fields were selected from each slide. *** p < 0.001, compared with the sham group. # p < 0.05, compared with the AMI group. &&&& p < 0.0001, compared with the AMI group.

Article Snippet: Paquinimod (Apexbio, Houston, TX, United States), also called ABR-215757 (a specific inhibitor of S100A9), binds to S100A9 in a Ca 2+ /Zn 2+ dependent way and blocks interaction with TLR4 ( ; ).

Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Immunostaining

Journal: Frontiers in Pharmacology

Article Title: Shuangxinfang Prevents S100A9-Induced Macrophage/Microglial Inflammation to Improve Cardiac Function and Depression-Like Behavior in Rats After Acute Myocardial Infarction

doi: 10.3389/fphar.2022.832590

Figure Lengend Snippet: Levels of S100A9, IL-1β, and TNF-α in the hippocampus and myocardium were determined by an ELISA assay. Values were expressed as mean ± SD. n = 6 per group. ** p < 0.01, **** p < 0.0001 compared with the sham group. # p < 0.05, ## p < 0.01, ### p < 0.001, compared with the AMI group. & p < 0.05, && p < 0.01, &&&& p < 0.0001, compared with the AMI group.

Article Snippet: Paquinimod (Apexbio, Houston, TX, United States), also called ABR-215757 (a specific inhibitor of S100A9), binds to S100A9 in a Ca 2+ /Zn 2+ dependent way and blocks interaction with TLR4 ( ; ).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Shuangxinfang Prevents S100A9-Induced Macrophage/Microglial Inflammation to Improve Cardiac Function and Depression-Like Behavior in Rats After Acute Myocardial Infarction

doi: 10.3389/fphar.2022.832590

Figure Lengend Snippet: PCF serum reduced the proinflammatory effect of recombinant protein S100A9 in BV2 cells via inhibiting TLR4. (A) Microglia cells were treated with protein S100A9 (0.01,0.02, 0.05, 0.1 μmol) for 6 h. CCK-8 assay was used to detect the viability of BV2 cells. (B) Effect of PCF serum with a concentration of 5%, 10%, and 20% on the viability of BV-2 cells. (C) Resting microglia (a, ×100 magnification; c, ×400 magnification) and activated microglia (b, ×100 magnification; d, ×400 magnification). Morphological changes of BV2 microglia after incubation with S100A9 protein for 6 h (b,d). (D) The ratio of the maximum radius to the minimum radius in BV2 microglia. (E) The level of S100A9, IL-1β, and TNF-α in the culture media was measured with ELISA. Each value represented the mean ± SD of three independent experiments. CS: control serum; CS-L: control serum low dose; CS-M: control serum medium dose; CS-H: control serum high dose; PCF: PCF serum; PCF-L: PCF serum low dose; PCF-M: PCF serum medium dose; PCF-H: PCF serum high dose; C34: TLR4 inhibitor; FA: ferulic acid; * p < 0.05, compared with the control group. # p < 0.05, compared with the S100A9 group. & p < 0.05, compared with the S100A9 group. ※ p < 0.05, compared with the S100A9 group.

Article Snippet: Paquinimod (Apexbio, Houston, TX, United States), also called ABR-215757 (a specific inhibitor of S100A9), binds to S100A9 in a Ca 2+ /Zn 2+ dependent way and blocks interaction with TLR4 ( ; ).

Techniques: Recombinant, CCK-8 Assay, Concentration Assay, Incubation, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Pharmacology

Article Title: Shuangxinfang Prevents S100A9-Induced Macrophage/Microglial Inflammation to Improve Cardiac Function and Depression-Like Behavior in Rats After Acute Myocardial Infarction

doi: 10.3389/fphar.2022.832590

Figure Lengend Snippet:

Article Snippet: Paquinimod (Apexbio, Houston, TX, United States), also called ABR-215757 (a specific inhibitor of S100A9), binds to S100A9 in a Ca 2+ /Zn 2+ dependent way and blocks interaction with TLR4 ( ; ).

Techniques: