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. 2011 Nov 1;9:186. doi: 10.1186/1479-5876-9-186

Figure 5.

Figure 5

Comparative expression analysis of EPO in CHO and Huh-7 cell line. (A) mRNA expression analysis showing equal level of expression of pcDNA3.1-K-SS-EPO vector in CHO and Huh-7 cell line. 1 μg of the total RNA extracted from CHO and Huh-7 cells after 24 hours post transfection was subjected to reverse transcriptase PCR. Lane 1, mock Huh-7 cells; Lane 2, EPO gene expression in Huh-7; Lane 3, CHO cells as mock; Lane 4, EPO gene expressing in CHO cell line. GAPDH is used as an internal control. (B) Comparative expression analysis of extra-cellular level of EPO protein through ELISA in CHO and Huh-7 cell line. 8 μg of the pcDNA3.1-K-SS-EPO vector was transfected in Huh-7 and CHO cell line, 24 hours post transfection media was collected and 200 μl of media was loaded on EPO antigen coated ELISA plates. Graph is showing that Huh-7 cells secreted the increased amount of EPO and difference in expression level is significant (P < 0.05). (C) Extra-cellular protein expression levels were determined by Western blot analysis in CHO and Huh-7 cell lines using EPO specific antibodies. Lane1, Commercially available purified EPO protein as standard; Lane 2, Huh-7 cells as mock; Lane 3, EPO expressed from Huh-7cell line; Lane 4, CHO cells as mock; Lane 5, EPO expressed from CHO cells.