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. 2019 Sep 18;18:533–545. doi: 10.1016/j.omtn.2019.08.027

Figure 2.

Figure 2

ADAMTS9-AS2 Is Required for Proper Chondrogenic Differentiation

(A) Targeting of ADAMTS9-AS2 in hMSCs by small hairpin RNA-constructed lentivirus led to significant depletion of the transcript, as measured by qPCR. (B) ADAMTS9-AS2 depletion affects cell condensation morphology and Alcian blue staining on cells on day 7. (C–E) Expression of chondrogenic genes measured by qPCR in ADAMTS9-AS2-depleted hMSCs after 14 days of chondrogenic differentiation in negative control and shRNA treatment group. (F) Synthetic glycosaminoglycan in ADAMTS9-AS2-knockdown hMSCs compared to negative controls on day 14, assessed by Safranine O staining, Alcian blue staining, and toluidine blue staining. Scale bar, 100 μm. (G) Antibody staining of paraffin-embedded sections of day 14 micromass showed Col2α1 and ACAN abundance between negative control and shRNA treatment group. Scale bar, 50 μm. (H and I) Quantitative statistics for dyeing results of (G) between negative control and shRNA treatment group. Experiments were performed in triplicate and error bars represent SD of a triplicate set of experiments. Data are shown as mean ± SD; *p, 0.05; **p, 0.01; ns, non-significant.