Journal: Journal of experimental & clinical cancer research : CR

Article Title: Regnase-1 downregulation promotes pancreatic cancer through myeloid-derived suppressor cell-mediated evasion of anticancer immunity.

doi: 10.1186/s13046-023-02831-w

Figure Lengend Snippet: Fig. 6 Suppression of CTLs by CD11b+ MDSCs is responsible for the acceleration of tumor progression by Regnase-1 downregulation. A-D Evaluation of phenotypes of orthotopic syngeneic tumors of WT or Regnase-1 KO murine pancreatic cancer cells. Representative macro images of pancreatic tumors (A). Relative mRNA levels of Cd8a, Ifng, Fasl, and Gzmb (B) (N = 6 per group). Representative images of HE (C, left panel) and CD8a immunostaining (C, right panel) and the number of CD8-positive cells (C, right) (N = 6 per group). Dot plots of CD3+CD8+ cells evaluated by flow cytometry (D, left) and the proportion of CD8 + cells among CD45+ cells (D, right) (N = 3 per group). E–H Evaluation of phenotypes of orthotopic syngeneic tumors of WT or Regnase-1-KO murine pancreatic cancer cells with or without depletion of CD8+ cells upon anti-CD8a antibody or IgG treatment. Experimental schematic (E). Dot plots of CD3+ and CD8.+ cells in WT or Regnase-1-KO syngeneic tumors upon anti-CD8a antibody or IgG treatment evaluated by flow cytometry (F). Tumor weights (G) (N = 6 per group). The relative mRNA levels of Cd8a, Ifng, Fasl, and Gzmb (H) (N = 6 per group). Student’s t test was used to evaluate differences between 2 groups. One-way ANOVA with Tukey’s post hoc test was used to compare differences among 4 groups. *P < 0.05, scale bars: 100 μm (insets)

Article Snippet: BE0061, a fully neutralizing mAb recognizing CD8a, and control IgG were obtained from Bioxcell.

Techniques: Immunostaining, Flow Cytometry

Journal: BJC reports

Article Title: A detoxified TLR4 agonist inhibits tumour growth and lung metastasis of osteosarcoma by promoting CD8+ cytotoxic lymphocyte infiltration.

doi: 10.1038/s44276-024-00120-3

Figure Lengend Snippet: Fig. 3 Lipo-MP-LPS induces immunological changes in both primary tumours and lung metastases. a, c Representative immunohis- tochemistry images of xenograft tumours and lung metastases stained with mouse specific anti-CD8a antibodies. Scale bar = 50 μm. b, d Quantification of CD8a positive cells in xenograft tumours and lung metastases. Data are represented as means ± SD; *p < 0.05, using Mann–Whitney U test. e, g Representative immunohistochemistry images of xenograft tumours and lung metastases stained with mouse specific anti-F4/80 antibodies. Scale bar = 50 μm. f, h Quantification of F4/80 positive cells in xenograft tumours and lung tissues. Data are represented as means ± SD; n = 6 mice per group, *p < 0.05, using Mann–Whitney U test. (i) iNOS, MHC II, TNF-α, CD206, and IL10 gene expression in xenograft tumours and lung metastases determined by RT-qPCR. Data are presented as means ± SD; n = 6 mice per group, *p < 0.05, **p < 0.01, using two-tailed t-test. NS not significant.

Article Snippet: In vivo depletion of CD8+ T cells Mice were depleted of CD8+ T cells by the intraperitoneal administration of an anti-CD8a antibody (YTS169.4, rat IgG2b; BioXCell, West Lebanon, NH, USA).

Techniques: Staining, MANN-WHITNEY, Immunohistochemistry, Gene Expression, Quantitative RT-PCR, Two Tailed Test

Journal: BJC reports

Article Title: A detoxified TLR4 agonist inhibits tumour growth and lung metastasis of osteosarcoma by promoting CD8+ cytotoxic lymphocyte infiltration.

doi: 10.1038/s44276-024-00120-3

Figure Lengend Snippet: Fig. 4 The tumour suppressive effect of Lipo-MP-LPS depends on CD8+ T cells. a Representative images of IHC staining for CD8a positive cells of xenograft tumours from mice treated with anti-CD8a antibody or IgG2b isotype. Scale bar = 100 μm. b CD8a expression in spleen tissues determined by RT-qPCR. Data are represented as means ± SD; n = 4. ***p < 0.001, using two-tailed t-test. c Tumour growth curve in C3H/HeN mice treated with empty liposome or Lipo-MP-LPS in addition to injection with anti-CD8a or isotype IgG2b antibodies (n = 6 mice per group). Data are shown as mean tumour volume ± SD. *p < 0.05, using Mann–Whitney U test. d Kaplan–Meier survival curves. p < 0.05 was considered significant using the log-rank test. Representative images of H&E staining of xenograft tumours (e) and lungs (g) for each group. Scale bar = 200 μm. f Quantification of necrotic areas in tumour tissues. h Quantification of lung metastasis areas. Data are presented as means ± SD; *p < 0.05, **p < 0.01; NS not significant, using Mann–Whitney U test.

Article Snippet: In vivo depletion of CD8+ T cells Mice were depleted of CD8+ T cells by the intraperitoneal administration of an anti-CD8a antibody (YTS169.4, rat IgG2b; BioXCell, West Lebanon, NH, USA).

Techniques: Immunohistochemistry, Expressing, Quantitative RT-PCR, Two Tailed Test, Injection, MANN-WHITNEY, Staining

Journal: BJC reports

Article Title: A detoxified TLR4 agonist inhibits tumour growth and lung metastasis of osteosarcoma by promoting CD8+ cytotoxic lymphocyte infiltration.

doi: 10.1038/s44276-024-00120-3

Figure Lengend Snippet: Fig. 6 Survival analysis of patients with osteosarcoma according to immune cell infiltration levels estimated by consensus TME. Of the 84 patients with osteosarcoma, the top third (28 patients) and bottom third (28 patients) were classified into high and low score groups, respectively. Kaplan–Meier curves of (a) CD8+ T cells (b) macrophage, (c) M1 macrophage, and (d) M2 macrophage infiltration for OS (left) and PFS (right) are presented. p < 0.05 was considered significant using the log-rank test.

Article Snippet: In vivo depletion of CD8+ T cells Mice were depleted of CD8+ T cells by the intraperitoneal administration of an anti-CD8a antibody (YTS169.4, rat IgG2b; BioXCell, West Lebanon, NH, USA).

Techniques:

Journal: Heliyon

Article Title: Aloe-emodin alleviates inflammatory bowel disease in mice by modulating intestinal microbiome homeostasis via the IL-4/IL-13 axis

doi: 10.1016/j.heliyon.2024.e34932

Figure Lengend Snippet: AE alleviates mice colitis induced by DSS. A : Changes in body weights of mice were monitored following treatment with different doses of AE in a mouse model of colitis induced by DSS. B : Intestinal inflammation marker LCN2 in feces. Levels of LCN2 in fecal samples collected from mice were measured as a marker of intestinal inflammation. C : H&E staining of colon tissue sections. Histological evaluation of colon tissue slices stained with H&E to assess tissue damage and inflammation. Representative images are shown. Data are shown as mean ± SD, ***P < 0.001: compared with control group; ##P < 0.01 and ###P < 0.001: compared with DSS group. Representative results from one out of three independent experiments.

Article Snippet: Additionally, fecal samples collected on day 15 were used for the detection of lipocalin-2 (LCN2) using an ELISA kit (RK-KOA0515; Rockland, Limerick, PA, USA).

Techniques: Marker, Staining, Control

Journal: Heliyon

Article Title: Aloe-emodin alleviates inflammatory bowel disease in mice by modulating intestinal microbiome homeostasis via the IL-4/IL-13 axis

doi: 10.1016/j.heliyon.2024.e34932

Figure Lengend Snippet: The content of LCN2 in control, DSS induced, and AE treated mice (ng/gfecal).

Article Snippet: Additionally, fecal samples collected on day 15 were used for the detection of lipocalin-2 (LCN2) using an ELISA kit (RK-KOA0515; Rockland, Limerick, PA, USA).

Techniques: Control

Journal: Nutrients

Article Title: Arctigenin from Saussurea medusa Maxim. Targets the PI3K/AKT Pathway to Inhibit Hepatocellular Carcinoma Proliferation and Induces Apoptosis

doi: 10.3390/nu17193151

Figure Lengend Snippet: Molecular Docking Results of ARC and core target proteins: ( a ) AURKA, ( b ) CCNB1, ( c ) CCND1, ( d ) CDK1, ( e ) CDK2, ( f ) CDK4, ( g ) GSK3B, ( h ) MAP2K1, ( i ) MTOR, ( j ) PIK3CA, ( k ) PIK3R1, ( l ) RAF1, ( m ) BRAF.

Article Snippet: Simvastatin reference compound (batch number 24240515002, purity >99%) were purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China); Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (batch number BST17383876), GSK3B antibody (batch number 24F107311F17), mechanistic target of rapamycin (mTOR) antibody (batch number 24F114106F24), Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) antibody (batch number BOS6238BP2883), pGSK3B (Ser9) antibody (batch number 24F087204A08) were all purchased from Wuhan Boster Biological Technology Co., Ltd. (Wuhan, China); metformin reference compound (batch number L396024, purity >97%) was purchased from Shanghai Haohong Biomedical Technology Co., Ltd. (Shanghai, China); LY294002 reference compound (batch number A825051337769) was purchased from APExBIO Technology LLC. (Houston, TX, USA); pPIK3CA (Tyr317) antibody (batch number 2i44771) and pmTOR (Ser248) antibody (batch number 8g6448) were purchased from Affinity Biosciences; Cyclophosphamide reference compound (batch number H32020857) was purchased from Jiangsu Hengrui Medicine Co., Ltd. (Lianyungang, China) Model 680 microplate reader, PowerPac Basic electrophoresis unit, and electrophoresis/transfer system were obtained from Bio-Rad (Hercules, CA, USA); The FlowSight flow cytometer was purchased from Cytek ® Biosciences Inc. (Fremont, CA, USA).

Techniques:

Journal: Nutrients

Article Title: Arctigenin from Saussurea medusa Maxim. Targets the PI3K/AKT Pathway to Inhibit Hepatocellular Carcinoma Proliferation and Induces Apoptosis

doi: 10.3390/nu17193151

Figure Lengend Snippet: Molecular Dynamics Simulation Results. ( a ) RMSD profiles of protein–ligand complexes over 50 ns simulations. ( b ) Binding energy profiles (enthalpy changes) of GSK3B and PIK3CA complexes.

Article Snippet: Simvastatin reference compound (batch number 24240515002, purity >99%) were purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China); Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (batch number BST17383876), GSK3B antibody (batch number 24F107311F17), mechanistic target of rapamycin (mTOR) antibody (batch number 24F114106F24), Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) antibody (batch number BOS6238BP2883), pGSK3B (Ser9) antibody (batch number 24F087204A08) were all purchased from Wuhan Boster Biological Technology Co., Ltd. (Wuhan, China); metformin reference compound (batch number L396024, purity >97%) was purchased from Shanghai Haohong Biomedical Technology Co., Ltd. (Shanghai, China); LY294002 reference compound (batch number A825051337769) was purchased from APExBIO Technology LLC. (Houston, TX, USA); pPIK3CA (Tyr317) antibody (batch number 2i44771) and pmTOR (Ser248) antibody (batch number 8g6448) were purchased from Affinity Biosciences; Cyclophosphamide reference compound (batch number H32020857) was purchased from Jiangsu Hengrui Medicine Co., Ltd. (Lianyungang, China) Model 680 microplate reader, PowerPac Basic electrophoresis unit, and electrophoresis/transfer system were obtained from Bio-Rad (Hercules, CA, USA); The FlowSight flow cytometer was purchased from Cytek ® Biosciences Inc. (Fremont, CA, USA).

Techniques: Binding Assay

Journal: Nutrients

Article Title: Arctigenin from Saussurea medusa Maxim. Targets the PI3K/AKT Pathway to Inhibit Hepatocellular Carcinoma Proliferation and Induces Apoptosis

doi: 10.3390/nu17193151

Figure Lengend Snippet: The Effect of ARC on GSK3B/PIK3CA/mTOR and their Phosphorylated Protein Expression. ( a , b ) Representative Western blotting bands of mTOR, pmTOR (Ser2481), PIK3CA, pPIK3CA (Tyr317), GSK3B, pGSK3B (Ser9), and GAPDH (loading control). ( c – k ) Quantitative analysis of the relative expression or phosphorylation ratio (phosphorylated/total protein) of these proteins in different treatment groups (Con, Met, Sim, LA, MA, HA, MA + LY). Note: Compared to the control group: * p < 0.05, # p < 0.0001.

Article Snippet: Simvastatin reference compound (batch number 24240515002, purity >99%) were purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China); Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (batch number BST17383876), GSK3B antibody (batch number 24F107311F17), mechanistic target of rapamycin (mTOR) antibody (batch number 24F114106F24), Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) antibody (batch number BOS6238BP2883), pGSK3B (Ser9) antibody (batch number 24F087204A08) were all purchased from Wuhan Boster Biological Technology Co., Ltd. (Wuhan, China); metformin reference compound (batch number L396024, purity >97%) was purchased from Shanghai Haohong Biomedical Technology Co., Ltd. (Shanghai, China); LY294002 reference compound (batch number A825051337769) was purchased from APExBIO Technology LLC. (Houston, TX, USA); pPIK3CA (Tyr317) antibody (batch number 2i44771) and pmTOR (Ser248) antibody (batch number 8g6448) were purchased from Affinity Biosciences; Cyclophosphamide reference compound (batch number H32020857) was purchased from Jiangsu Hengrui Medicine Co., Ltd. (Lianyungang, China) Model 680 microplate reader, PowerPac Basic electrophoresis unit, and electrophoresis/transfer system were obtained from Bio-Rad (Hercules, CA, USA); The FlowSight flow cytometer was purchased from Cytek ® Biosciences Inc. (Fremont, CA, USA).

Techniques: Expressing, Western Blot, Control, Phospho-proteomics