Journal: International journal of molecular sciences

Article Title: TGF-β Type I Receptor Signaling in Melanoma Liver Metastases Increases Metastatic Outgrowth.

doi: 10.3390/ijms24108676

Figure Lengend Snippet: Figure 6. Secretome analysis of B16F10 caALK5 cells. (A) Mass spectrometry analysis of the secretome of control and caALK5 expressing cells. Cells were labeled with azidohomoalanine and treated with control or dox (1 µg/mL) for 48 h prior to biotin labeling and pulldown, followed by mass spectrometry analysis. Significantly upregulated (green) and downregulated (red) proteins in caALK5 expressing B16F10 secretome are shown. (B) Proteins upregulated in caALK5 expressing B16F10 cells were analyzed for enrichment in GO annotation and KEGG pathways using Metascape. Top 20 significantly upregulated processes are shown, colored by p values. (C) Analysis of CD3 staining in the tumor border and center. All metastases per group are shown, from n = 10 in the control and n = 70 in the dox-treated group. Significance was calculated using two-way ANOVA, * p ≤0.05.

Article Snippet: The following primary antibodies were used: platelet-derived growth factor receptor (PDGFR)-β (1/100, #31695, Cell Signaling Technology, Leiden, The Netherlands), CD3 (1/100, #NB600–1441SS, Novus Bio, Abingdon, UK), CD8 (1/100, #14- 0808-82, Invitrogen, Bleiswijk, The Netherlands), plasminogen activator inhibitor (PAI)-1 (1/ab222754, Abcam), ionized calcium-binding adapter molecule1 (Iba-1) (1/1000, #019- 19741, Fujifilm, Dusseldorf, The Netherlands), C-C chemokine receptor type 2 (CCR2) (1/100, ab273050, Abcam, Amsterdam, The Netherlands), and C-type lectin domain family 4 (CLEC4F) (PA5-47396, Thermofisher, Bleiswijk, The Nethelands).

Techniques: Mass Spectrometry, Control, Expressing, Labeling, Staining

Journal: World Journal of Oncology

Article Title: Sphingosine-1-Phosphate Inhibition Increases Endoplasmic Reticulum Stress to Enhance Oxaliplatin Sensitivity in Pancreatic Cancer

doi: 10.14740/wjon1768

Figure Lengend Snippet: SPHK2-S1P inhibition enhances antitumor activity of Oxa in an orthotopic KPC model. Following treatment for 21 days, orthotopic tumors were harvested and analyzed. (a) In vivo experimental schema. After orthotopic injection of KPC cells, mice were randomized to four treatment groups of 7 - 10 mice each, starting 7 days post-operatively. Oxa at 3 mg/kg (red arrows) was intraperitoneally injected twice weekly and 50 mg/kg of ABC (green arrows) was administrated by oral gavage three times per week. Tumors were harvested 3 weeks after tumor implantation. (b) Analysis of harvested tumor weights by treatment group (*P < 0.05, **P < 0.01). (c) Analysis of IHC stained tumor samples of positively stained nuclei/HPF for Ki67, CC3, CD3, and CD8 between treatment groups. Intensity of HMGB1 staining was quantified by calculating an H-score (*P < 0.05, **P < 0.01). (d) Representative images of IHC stained images of tumors in different treatment groups. ABC: ABC294640; HPF: high power field; IHC: immunohistochemical; Oxa: oxaliplatin; S1P: sphingosine-1-phosphate; SPHK2: sphingosine kinase 2.

Article Snippet: Sections were also stained with Ki67 antibody (1:400 dilution, cat#12202, Cell Signaling), cleaved caspase-3 (CC3) antibody (1:400 dilution, cat#9661, Cell Signaling), HMGB1 antibody (1:1,000 dilution, cat#6893, Cell Signaling), CD3 antibody (1:400 dilution, cat# NB600-1441SS, Novus Biologicals), and CD8 antibody (1:400 dilution, cat#98941s, Cell Signaling).

Techniques: Inhibition, Activity Assay, In Vivo, Injection, Tumor Implantation, Staining, Immunohistochemical staining