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. Author manuscript; available in PMC: 2014 Aug 27.
Published in final edited form as: J Immunother. 2009 Feb-Mar;32(2):145–156. doi: 10.1097/CJI.0b013e318193d31e

FIGURE 1.

FIGURE 1

Construction and characterization of Ad-RANTES-E1A and Ad-E1A vectors. A, Schematic representation of pShuttle vectors is shown. IRES indicates internal ribosome entry site; mRANTES, mouse regulated upon activation, normally T expressed, and presumably secreted. B, The integrity of the RANTES, IRES, and E1A in the produced adenovirus was verified by PCR on viral DNA template. Lane 1, PCR product of mRANTES (276 bp); lane 2, PCR product for IRES (610 bp); lane 3, PCR product for E1A (2100 bp); lanes 4 and 5, 100 bp ladder and 1 kb ladder (New England Biolabs), respectively. C, In vitro expression of RANTES. Western blot analysis of cell extracts from JC cells infected with Ad-RANTES-E1A (lane 1) and Ad-E1A (lane 2). Western blots were performed as described in Materials and Methods. Molecular weight in kDa is indicated on the right and the RANTES band (~7.9 kDa) is indicated by the arrow on the left. D, Secretion of RANTES from the transduced cells by the recombinant adenoviruses was quantitated by ELISA in the supernatants of 293 HEK and JC tumor cells. ELISA indicates enzyme-linked immunosorbent assay; HEK, human embryonic kidney cell; PCR, polymerase chain reaction.