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. 2019 Dec 5;61(3):432–444. doi: 10.1194/jlr.M094540

Fig. 3.

Fig. 3.

Western blots of cell lysates and immunoprecipitated culture media from HEK293 cells transfected with WT and R1771C apo(a)-GFP vectors. HEK293 (A) and Huh7 (B) cells were seeded in 6-well plates and either mock-transfected or transfected with WT or R1771C apo(a)-GFP vectors. At 72 h posttransfection, cell lysates and media were harvested. apo(a) was isolated from culture media by immunoprecipitation on Dynabeads protein G with an anti-Lp(a) antibody and eluted under reducing conditions. Cell lysates and immunoprecipitants (IPs) were run on SDS 4% polyacrylamide gels. Plasma from an individual with apo(a) KIV isoform size of 18 was run as a positive control on lysate gels. After transfer to nitrocellulose, membranes were probed with a goat anti-Lp(a) antibody and a HRP-labeled anti-goat IgG secondary antibody. For loading controls, lysates or immunoprecipitants were run on SDS 10% polyacrylamide gels, transferred to nitrocellulose, and probed with either an anti-actin antibody for lysates or an HRP-labeled anti-goat IgG antibody for immunoprecipitants. Gels were loaded with molecular mass standard labeled in kilodaltons. HMM, high molecular mass; LMM, low molecular mass.