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. 2021 May 26;80(9):1209–1219. doi: 10.1136/annrheumdis-2021-219969

Figure 2.

Figure 2

Targeting circPDE4B expression affects matrix-degrading and anabolic factors in human chondrocytes (HCs). (A) circPDE4B expression in HCs transfected with circPDE4B siRNAs or negative control siRNA (n=9, 3 donors for three replicates); *p≤0.05. (B) Viability of HCs infected with circPDE4B shRNA #2/#3 adenovirus or control shRNA adenovirus (n=9, 3 donors for three replicates); *p≤0.05. (C, D) mRNA and protein levels of MMP3, MMP13, ADAMTS4, COL2A1 (or COL2 protein), SOX9 and aggrecan in HCs infected with circPDE4B shRNA #2/#3 adenovirus or control shRNA adenovirus (n=9, 3 donors for three replicates); *p≤0.05. (E) Immunofluorescence analysis of MMP3, MMP13, COL2 and aggrecan. (F) Quantification of immunofluorescence analysis (n=9, 3 donors for three replicates); *p≤0.05. (G) Alcian blue staining of shRNA-treated HCs. (H) Viability of HCs infected with a circPDE4B overexpression adenovirus or control adenovirus (n=9, 3 donors for three replicates); *p≤0.05. (I, J) mRNA and protein levels of MMP3, MMP13, ADAMTS4, COL2A1 (or COL2 protein), SOX9 and aggrecan (n=9, 3 donors for three replicates); *p≤0.05. (K) Immunofluorescence of MMP3, MMP13, COL2 and aggrecan. (L) Quantification of immunofluorescence analysis (n=9, 3 donors for three replicates); *p≤0.05. (M) Alcian blue staining of HCs treated with IL-1β with or without circPDE4B overexpression. IL-1β, interleukin-1β; NC, negative control.