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Quantifying PCSK9 Secretion in Human HuH7 Cells Transfected with Different Constructs

ELISA demonstrates the linearity of WT PCSK9 secretion from HuH7 cells transfected with WT PCSK9 untagged (WT.NT) and V5-tagged WT PCSK9 (WT.V5), but no secretion with Q152H PCSK9 (QH.V5), with amounts of cDNA as shown, n=3, ave ± SEM. HuH7 cells were grown at 37°C in DMEM + 10% FBS + gentamycin (28 µg/ml). Cells (3x10^5) were transiently transfected with cDNA encoding human PCSK9 (pIRES2) using Effectene protocol from Qiagen 24 hours following plating.

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Quantifying PCSK9 Secretion in Human HuH7 Cells Transfected with Different Constructs

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  1. 350 300 250 200 Secreted PCSK9 (ng/ml) 150 100 50 0 Input DNA WT NT WT V5 QH V5 ng Input DNA 125 250 500 125 250 500 125 250 500 250 ng non-relevant DNA 375 250 0 375 0 375 250 0 Supplemental Figure 1 ELISA demostrating the linearity of WT PCSK9 Secretion from HuH7 cells transfected with WT PCSK9 untagged (WT NT) and V5-tagged WT PCSK9 (WT V5) but no secretion with Q152H PCSK9 (QH V5), with amounts of cDNA as shown. n=3; ave  SEM. HuH7 cells were grown at 37C in DMEM + 10% FBS + gentamycin (28 g/ml). Cells (3X105) were transiently-transfected with cDNA encoding human PCSK9 (pIRES2) using Effectene protocol from Qiagen 24 hours following plating.

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