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. 2017 Dec 15;144(24):4510–4521. doi: 10.1242/dev.152504

Fig. 4.

Fig. 4.

Effect of lncRNA depletion on MSC chondrogenic gene expression. (A) MSCs were transfected for 2 days with SOX9-AS1 or ROCR-targeting or non-targeting control (siCon) siRNA prior to chondrogenic differentiation in hanging transwell inserts. RNA was extracted and expression of the indicated genes measured by real-time RT-PCR. (B-D) MSCs were transfected for 1 day with SOX9-AS1 or ROCR-targeting or non-targeting control siRNA prior to chondrogenic differentiation in a V-bottom 96-well plate for up to 24 h. RNA and protein was extracted and expression of SOX9 mRNA (B) or protein (C) measured by real-time RT-PCR or immunoblotting, respectively. (D) Expression of L-SOX5a and SOX6. Values are mean±s.e.m. of data pooled from three (A) or four (B-D) MSC donors. *P<0.05; **P<0.01; ***P<0.001 for lncRNA siRNA versus non-targeting siRNA. Significant differences between sample groups were assessed by one-way analysis of variance followed by the Bonferroni post-hoc test for multiple comparisons or a two-tailed Student's t-test was performed for single comparisons.