Supplemental Figure 1. from VEGF Potentiates GD3-Mediated Immunosuppression by Human Ovarian Cancer Cells

Autor: Tonya J. Webb, Jonathan P. Schneck, Mathias Oelke, Robert L. Giuntoli, Simeon E. Goldblum, Sarah Spiegel, Sarah M. Temkin, Irina V. Tiper
Rok vydání: 2023
DOI: 10.1158/1078-0432.22455875.v1
Popis: Ovarian cancer associated ascites contains soluble inhibitory factors. The ascites was cleared of cellular debris by centrifugation as previously described (18). (A, B) The ascites was filtered 0.2 micron filter and fractionated- f1: 50. (C) The ascites was treated with 50 μg/ml proteinase K at 37{degree sign}C for 1 hour and then incubated at 95{degree sign}C for 10min to heat inactivate the enzyme. The antigen presenting cells were treated with the clarified supernantants for 4 h at 37{degree sign}C, then washed extensively with PBS, and cocultured with NKT hybridomas for 20-24 h at 37{degree sign}C. Cytokine released (IL-2) was measured as an indication of NKT cell activation and was measured by standard sandwich ELISA.
Databáze: OpenAIRE