Supplemental Figure S16 from VAX014 Activates Tumor-Intrinsic STING and RIG-I to Promote the Development of Antitumor Immunity
Year: 2026
Type: supplementary-materials
Abstract: Supplemental Figure S16. In vitro characterization of all MB49 Double KO clones. A) RIG-I expression in MB49WT (WT) vs. MB49STING/RIG-I KO (Double KO) cells analyzed by Western blot. GAPDH was run as a loading control (n= 1). B) STING expression in MB49 WT vs. MB49 Double KO cells analyzed via Western blot, using Vinculin as a loading control (n= 1). C) MB49 WT, MB49 single KO (STING KO or RIG-I KO), and MB49 Double KO cells were plated (5,000 cells/well) and treated with a titration (MOI 2,000:1 to 4:1) of VAX014 rBMCs for 24 hours and assayed for cell killing. EC50 values calculated by GraphPad Prism 10 are shown (n= 2-3). D) Surface expression of PD-L1 in MB49 Double KO clones as determined by flow cytometry (n= 3). Cells were plated (5.0x105 cells/well) and treated with DMXAA (30 μg/mL) or VAX014 rBMCs (MOI 300) for ∼20 hours. E) IFN-β content in supernatants from D) as measured via ELISA (n= 2). F) Surface expression of PD-L1 and MHC-I in MB49 single KO (STING KO or RIG-I KO) and MB49 Double KO cells co-incubated with VAX014-treated MB49 WT supernatants for ∼20 hours, analyzed by flow cytometry (n= 1). Histograms depict mean ± SEM of all combined experiments and dashed lines represent the baseline expression value of the corresponding untreated control. SRKO=Double KO cell line where RIG-I was ablated from MB49 STING KO cells; RSKO=Double KO cell line where STING was ablated from MB49 RIG-I KO cells. ns = not significant, all compared to STING KO or RIG-I KO. #indicates which Double KO clone is represented in Figure 4.
Language: English
Subfield: Immunology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W7165056392