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Figure 3 | Arthritis Research & Therapy

Figure 3

From: MicroRNA-33a regulates cholesterol synthesis and cholesterol efflux-related genes in osteoarthritic chondrocytes

Figure 3

ΜicroRNA-33a affects Smad7 expression in human chondrocytes. (A) TargetScan 6.2 and miRanda identified one conserved sequence in the 3′ untranslated region of human Smad7 mRNA that was completely complementary to microRNA (miR)-33a. (B) Relative expression levels of miR-33a in normal human chondrocytes (n = 3 from 3 different donors) transfected with miR-33a mimic or negative control. U6 was used for normalization of the real-time PCR data. The data are expressed as mean and standard error of the mean (SEM) of three independent experiments, each of which was run in duplicate. *P < 0.05 versus negative control. (C) Relative expression levels of Smad7 mRNA 24 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 SEM of three independent experiments, each of which was run in triplicate. *P < 0.05 versus negative control. (D) Relative expression levels of miR-33a in human osteoarthritis (OA) chondrocytes transfected with miR-33a inhibitor or negative control (n = 3 from 3 different donors). U6 was used for normalization of the real-time PCR data. The data are expressed as mean and SEM of three independent experiments, each of which was run in duplicate. *P < 0.05 versus negative control. (E) Relative expression levels of Smad7 mRNA 24 hours after transfection of 50 nM anti-miR-33a or negative control in human OA 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 three independent experiments, each of which was run in triplicate. *P < 0.05 versus negative control.

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