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Fig. 3 | BMC Biotechnology

Fig. 3

From: Stable high-level expression of factor VIII in Chinese hamster ovary cells in improved elongation factor-1 alpha-based system

Fig. 3

Integrity of the FVIII’s ORF in the genome DNA and mRNA of the 11A4H cell line and Southern blot analysis of genomic DNA. Panel a – PCR with the annealing temperature gradient for genomic DNA from 11A4H cell line. Panel b – PCR for cDNA, prepared from 11A4H cell line and control un-transfected CHO DG44 cells. Panel c – Southern blot analysis of genomic DNA from 11A4H and 21A7B cell lines and from un-transfected control. “FVIII probe” - probe toward FVIII ORF, “IRES-DHFR-bla” – probe toward corresponding expression vector areas. Contrast for scans of developed membranes was enhanced to add visibility. Red triangles depict annealing temperature gradients of 53 °C–68 °C for all reactions, DNA fragments sizes in bp. Expected size of the correct PCR product – 4818 bp. Images of the agarose gels used for blotting are presented on the Additional file 4: Figure S3

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