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. 2019 Mar 19;10(23):2252–2269. doi: 10.18632/oncotarget.26785

Figure 1. Schematic diagram of RRV-scFv-PDL1 plasmid DNAs.

Figure 1

(A) Two pairs of single-chain variable fragment (scFv) against PD-L1 were encoded in pAC3 RRV backbone. One pair consists of scFv with and without the Fc from human IgG1, designated as pAC3-scFv-PDL1 and pAC3-scFvFc-PDL1, respectively. Another pair consists of scFv-PDL1 and scFvFc-PDL1 with HA and Flag epitope incorporated at the C-terminus, designated as pAC3scFvHF-PDL1, pAC3-scFvFc-HF-PDL1. Filled rectangle indicates leader sequence derived from human IL-2. (B) Western blot analysis of viral envelope proteins produced transient transfection in 293T cells. Twenty micrograms of total protein lysates were loaded per well. Membranes were incubated (left panel) with anti-HA and anti-Flag antibody which detects HA- and Flag-tagged scFv-PD-L1 and scFvFc-PD-L1, respectively, or (right panel) with anti-2A peptide antibody which detects Env-scFv polyprotein (Env-scFv), unprocessed viral precursor envelop protein separated from the Env-scFv polyprotein (Env-2A), and processed viral envelop protein tagged with the 2A peptide at the C-terminus (p15E-2A). Anti-GAPDH antibody (lower left panel) was included as loading control.