Fig. 2
The expression of CB6 with VEEV-VRP delivery in vitro. a Schematic representation of the packaging system of VEEV-VRP-CB6 including the VEEV-rep-CB6, and two helper RNAs as described previously,32 and the transduction processes of BHK-21 cells with VEEV-VRP-CB6. b BHK-21 cells were infected with VEEV-VRP-CB6 at different MOIs (0.1/1/10), the cells were fixed and stained for the expression of CB6 mAb (red) and VEEV-nsP1 (green) using Alexa FluorTM 568 conjugated goat anti-human IgG (H + L) and anti-VEEV nsP1 mouse serum respectively at 24 h post-infection (hpi). The mock-infected BHK-21 cells were used as a negative control. Nuclei were stained with DAPI (blue). Scale bars represent 50 μm. c Quantification of CB6 expression levels by antigen (RBD)-specific ELISA using the supernatants collected at 24 hpi from the VEEV-VRP-CB6 infected BHK cells with different MOIs (0.1/1/10) and mock treated BHK-21 cells. The antibody titers were determined in triplicates. d The VEEV-VRP-CB6 infected BHK-21 cells (MOI = 1) and the mock treated cells were lysed with RIPA at 48 hpi and subjected to western blotting using the HRP conjugated goat anti-human IgG (H + L) and the anti-β-actin antibody. The expression of β-actin was used as an internal control
