Figure 7: LPA mRNA and Protein Expression Were Detected in Primarily Cultured HAVICs and Aortic Valve Tissues

(A) RT-PCR detection of LPA mRNA expression in primarily cultured HAVICs. Lane 0: DNA marker; lanes 2, 5, and 8 show positive signal at products 136 bp. (B) RT-PCR detection of LPA mRNA expression in stenotic aortic valves. Lane 0: DNA marker; lanes 1, 2, 5, 6, 7, 9, and 10 show positive signal at 136 bp. (C) Sequencing of gel purified RT-PCR product confirms the correct identity of LPA mRNA. (D) LPA protein expression in stenotic aortic valve tissues and HAVICs. (Top) Lanes 1 through 3, nonstenotic valves; lanes 4 through 11, stenotic valves; lane 12, human serum sample serving as loading marker. Lanes 8 and 9 show double bands indicating possible dual isoforms. (Bottom) LPA protein expression in 7 cultured HAVICs. Lane 1 show double bands indicating possible dual isoforms. (E) Immunofluorescence cytochemistry detection of LPA in HAVICs using mouse anti-LPA antibody and Alexa Fluor 488 donkey anti-mouse immunoglobulin G (green); solid arrows show various-sized cellular vesicles. Red indicates pseudocolored F-actin stained with ActinRed reagent; blue indicates nuclei stained with NucBlue reagent. LPA = apolipoprotein(a) gene; mRNA = messenger ribonucleic acid; RT-PCR = real-time polymerase chain reaction; other abbreviations as in Figure 1.