Figure 1: Effects of Lp(a) Treatment on HAVICs

HAVICs were incubated in OSM in the presence or absence of 50 μg/ml purified Lp(a) (Athens Research & Technology, Inc., Athens, Georgia) for 48 h. (A) Cell proliferation measured using an MTT protocol and presented as relative cell proliferation rate. *p = 0.013 vs. OSM control. (B) Medium and cellular ALP activity measured with an ALP activity colorimetric assay kit and normalized to total protein concentration. Lp(a)-treated cells showed higher ALP activity in medium (**p = 0.0001) and cell lysate (*p = 0.003) than OSM control. (C) Medium phosphate release measured using a phosphate colorimetric assay kit (BioVision, Inc., Milpitas, California). *p = 0.0001 vs. OSM (control). (D) No detectable OxPLs were seen on purified Lp(a) using anti-OxPL E06 antibody. Purified human Lp(a) (1 μg) was loaded on 6% sodium dodecyl sulfate polyacrylamide gel electrophoresis. Lane 1: Lp(a) was detected using anti-Lp(a) antibody (diluted at 1:10,000; Novus Biologicals, LLC, Littleton, Colorado); lane 2: no OxPLs were detected in purified Lp(a) using E06 antibody (diluted at 1:1,000; Avanti Polar Lipids, Inc., Alabaster, Alabama). ALP = alkaline phosphatase; HAVICS = human aortic valve interstitial cells; Lp(a) = lipoprotein(a); OSM = osteogenic medium; OxPL = oxidized phospholipid.