Figure 5.

Atg2a was involved in miR-378-regulated inflammatory response and autophagy of chondrocytes

(A) Conservation of the miR-378-3p-binding site on Atg2a 3′ UTR (shaded region) among different species. The wild-type (WT) and mutation (mut) forms of Atg2a 3′ UTR luciferase reporter vector are shown. (B) Effects of miR-378-3p on the luciferase activity of pmiRGLO vectors incorporated with Atg2a-wt or Atg2a-mut sequence were measured (n = 6; ∗∗∗p < 0.001). (C) The mRNA and protein expression level of Atg2a in WT and miR-378 chondrocytes were measured respectively (n = 3; ∗p < 0.05). (D–H) The mRNA expression levels of iNos (D), Cox2 (E), Col2a1 (F), Mmp13 (G), and Beclin-1 (H) of WT and miR-378 chondrocytes upon Atg2a overexpression under inflammatory conditions were assessed by real-time PCR (n = 6; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001). (I and J) The protein expression level of iNos, Cox2, Col II and Mmp13 (I) as well as Beclin-1 and LC3 (J) of WT and miR-378 chondrocytes upon Atg2a overexpression under inflammatory conditions were determined by western blot analysis. (K) Fluorescence images of Cyto-ID dye-stained WT and miR-378 chondrocytes upon Atg2a overexpression under inflammation conditions. Scale bar, 100 μm.