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. 2014 Nov 26;6(2):1079–1089. doi: 10.18632/oncotarget.2835

Figure 1. Identification of adenovirus vector in HCC cell lines.

Figure 1

(A) Schematic diagram of the oncolytic adenoviruses and amplification of the early replication gene E1a as well as the therapeutic gene Hsp70 was performed by PCR in adenoviral vectors AdSurp-Hsp70 and AdSurp-EGFP. The Survivin promoter was used to regulate the adenoviral E1a gene, and the expression cassettes of Hsp70 and EGFP were inserted into the upstream of E1a gene, then generated the recombinant oncolytic adenoviruses AdSurp-Hsp70 and AdSurp-EGFP. ITR: inverted terminal repeats; ψ: adenovirus 5 packaging signal; CMV: cytomegalovirus promoter; Surp: Survivin promoter. (B) The indicated cell lines were seeded into 24-well plates at a concentration of 1×105 cells/well, and infected with AdSurp-EGFP at an MOI of 1 pfu/cell, cultured for 48 h and observed the EGFP-positive cells under a fluorescence microscope, original magnification: ×200. (C) The cell lines were seeded into 24-well plates at a concentration of 1×105 cells/well, and infected with AdSurp-Hsp70 at an MOI of 1 pfu/cell, cultured for 48 h and the expression of Hsp70 was examined by Western blotting; ***P<0.001.