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0 50 100 150 200 250 300 350 Secreted PCSK9 (ng/ml) WT V5 QH V5 ng Input DNA 125 250 ng non-relevant DNA 375 250 0 375 250 0 WT NT 375 250 0 Input DNA 500 125 250 500 125 250 500 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 37C in DMEM + 10% FBS + gentamycin (28 g/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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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. - PowerPoint PPT Presentation
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Se
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PC
SK
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ng
/ml)
WT V5 QH V5
ng Input DNA 125 250
ng non-relevant DNA375 250 0 375 250 0
WT NT
375 250 0
Input DNA
500 125 250 500 125 250 500
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 37C 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.