Figure 4

ΜicroRNA-33a affects ATP-binding cassette transporter A1 and apolipoprotein A1 expression in human chondrocytes. (A) TargetScan 6.2, miRanda and miRDB identified three conserved sequences in the 3′ untranslated region of human ATP-binding cassette transporter A1 (ABCA1) mRNA that were completely complementary to microRNA (miR)-33a. (B) and (C) Relative expression levels of ABCA1 (B) and apolipoprotein A1 (ApoA1) (C) mRNA at 6, 24 and 48 hours after transfection of 30 nM miR-33a or negative control in normal human chondrocytes (n = 6 from 6 different donors). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used for normalization of the real-time PCR data. The data are expressed as mean and standard error of the mean (SEM) of two independent experiments, each of which was run in triplicate. *P < 0.05 versus negative control. (D) Relative expression levels of matrix metalloproteinase (MMP)-13 mRNA at 6, 24 and 48 hours after transfection of 30 nM miR-33a or negative control in normal human chondrocytes (n = 4 from 4 different donors). GAPDH was used for normalization of the real-time PCR data. The data are expressed as mean and SEM of two independent experiments, each of which was run in triplicate. *P < 0.05 versus negative control. (E) and (F) Relative expression levels of liver X receptor α (LXRα) (E) and LXRβ (F) mRNA at 6, 24 and 48 hours after transfection of 30 nM miR-33a or negative control in normal human chondrocytes (n = 4 from 4 different donors). GAPDH was used for normalization of the real-time PCR data. The data are expressed as mean and SEM of two independent experiments, each of which was run in triplicate. NS, Not significant.