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Fig. 3 | BMC Cancer

Fig. 3

From: Spatial and temporal epithelial ovarian cancer cell heterogeneity impacts Maraba virus oncolytic potential

Fig. 3

Differences in LDLR expression can impact MRBV entry. a. iOvCa147-F8 and iOvCa147-G4 cells were seeded at 75,000 cells per well of a 24-well plate. After 24 h, cells were infected with MRBV at an MOI of 1 for 1 h. Supernatant was collected and non-cell associated virus was titrated using Vero cells; supernatant from infections containing no cells were used as a control for total uninfected virus. b. Western blot were performed using lysates collected from iOvCa147-F8 and iOvCa147-G4 cells infected with MRBV at an MOI of 1, or UV-inactivated MRBV and no virus as controls. Actin served as a loading control. The graph represents quantification of LDLR expression performed using Bio-Rad Image Lab software and normalized to actin. c. Two different MRBV-sensitive subclones iOvCa147-F8 and iOvCa147-E2 cells were seeded at 20,000 cells per well of a 48-well plate. Cells were transfected with siNT or siLDLR and 48 h post-transfection, cells were harvested for protein lysates. Western blot for LDLR was performed and actin was used as a loading control. d. iOvCa147- F8 and iOvCa147-E2 cells transfected with siNT or siLDLR were treated with 100 nM of RAP for 1 h at 48 h post-transfection followed by MRBV infection for another 1 h. media was collected for titration of MRBV virus on Vero cells via plaque assay; treatments containing no cells were performed for normalization. e. iOvCa147- F8 and iOvCa147-E2 cells transfected with siNT or siLDLR were treated with 100 nM of RAP for 1 h at 48 h post-transfection followed by MRBV infection for another 1 h. Cells were assayed for viability using CellTiter-Glo at 72 h post-infection. (*, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001, as determined by one-way ANOVA and Tukey’s posthoc test)

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